Effect of acute exposure to electronic cigarette on bronchoalveolar lavage fluid and pulmonary surfactant protein in mice / 预防医学

Objective@#To assess the effects of acute exposure to electronic cigarette ( e-cigarette ) on leukocyte and total protein levels in bronchoalveolar lavage fluid ( BALF ) and pulmonary surfactant protein expression in a mouse model, so as to provide insights into the elucidation of the mechanism underlying the damages to the respiratory system caused by e-cigarette.@*Methods@#Twenty-one C57BL/6N female mice were randomly divided into the blank control group, the solvent control group and the nicotine group. Mice in the solvent control group and the nicotine group were exposed to the solvent aerosol or e-cigarette aerosol containing 25 mg/mL nicotine for 3 hours daily, while mice in the blank control group were bred in clean air. Following 3-day exposure, mouse BALF and lung specimens were collected. The cell morphology was observed using microscopy following Wright-Giemsa staining and the leukocyte count was estimated in BALF, while the total protein expression was quantified using bicinchoninic acid ( BCA ) assay. In addition, the mRNA expression of pulmonary surfactant protein genes was detected in mouse lung specimens using quantitative real-time PCR ( qPCR ) assay.@*Results@#All mice in three groups grew well without obvious abnormality or death seen. Wright-Giemsa staining showed a higher number of mononuclear macrophages in mouse BALF in the nicotine group than in the blank control group and the solvent control group. The leukocyte counts were ( 2.00±0.77 )×107, ( 1.79±0.99 )×107 and ( 4.00±1.35 )×107 cells/L ( F=9.199, P=0.002 ), and the total protein levels were ( 0.16±0.03 ), ( 0.12±0.02 ) and ( 0.16±0.04 ) mg/mL in mouse BALF in the blank control group, solvent control group and nicotine group ( F=3.610, P=0.048 ), and the relative mRNA expression of pulmonary surfactant protein B (SP-B) and SP-D was 1.00±0.14, 0.82±0.12 and 0.74±0.07 ( F=5.491, P=0.028 ), and 1.00±0.06, 0.90±0.02 and 0.71±0.15 in mouse lung specimens, respectively ( F=10.460, P=0.005 ). The leukocyte count was significantly higher in the nicotine group than in the blank control group and solvent control group (P=0.007, 0.003), and the total protein content was higher in the nicotine group than in the solvent control group ( P=0.060 ), while the relative SP-B mRNA expression was lower in the nicotine group than in the blank control group ( P=0.025 ), and the relative SP-D mRNA expression was lower in the nicotine group than in the blank control group and solvent control group ( P=0.004, 0.041 ).@*Conclusion@#Acute exposure to e-cigarette results in elevated intrapulmonary inflammatory responses, pulmonary capillary barrier impairment and reduced pulmonary surfactant protein expression.

Clinical significance of Clara cell secreted protein 16 and pulmonary surfactant protein-A in early diagnosis of neonatal acute respiratory distress syndrome / 中国小儿急救医学

Objective:To investigate the clinical significance of changes of serum Clara cell secretory protein(CC16) and pulmonary surfactant protein A(SP-A) in neonates with acute respiratory distress syndrome(ARDS).Methods:The data of 30 neonates with ARDS who needed mechanical ventilation in neonatal intensive care unit of Xi′an Children′s Hospital from January 2016 to November 2018 were collected as observation group, including 12 cases in mild group, 10 cases in moderate group and 8 cases in severe group.The data of healthy newborns during the same period were taken as control group.The serum levels of CC16 and SP-A were detected by ELISA.The serum levels of CC16 and SP-A among different groups were compared.Results:The levels of serum CC16 and SP-A in ARDS group were (59.35±3.67)mg/L and(75.38±6.27)mg/L respectively, (11.26±1.32)mg/L and(18.15±2.69)mg/L in healthy group.The difference was significant( P<0.05). And the differences of serum CC16 and SP-A levels among different degree ARDS groups were significant( P<0.05). The levels of serum CC16 in mild, moderate and severe subgroup were(38.27±16.01)mg/L, (51.25±15.63)mg/L, (84.76±13.12)mg/L and SP-A were(47.02±7.18)mg/L, (73.12±7.98)mg/L, (96.45±12.50)mg/L, which increased with disease severity. Conclusion:Serum CC16 and SP-A are increased and correlated with the severity of neonatal ARDS, which may be used as the index for evaluating the severity of neonatal ARDS in the future.

Progress on PM2.5 and its adverse impact on lung tissues / 中华实用儿科临床杂志

The atmospheric particulate matter(PM) is widely regarded as one of major environmentally and unfriendly ambient air pollution, and therein PM2.5 (diameter≤2.5 μm) is most closely related to human health.Because of its smaller diameter with longer suspension duration, PM can absorb many pathogenic microorganisms, and easily enter into the deep of airway and then deposit on the bronchus and alveoli, and it brings damage to the lung tissues and the surfactant proteins.PM can give rise to oxidative stress, inflammation response, cells and DNA damage.Now, this review focuses on the characterization and composition of PM, as well as the impact of PM2.5 on the lung, surfactant proteins and human health, which helps to call for more people to pay attention to this environmental issue in order to better mitigate and prevent the damage caused by PM.

Diagnostic value of lung injury biomarkers in neonatal acute respiratory distress syndrome / 国际儿科学杂志

Objective@#To investigate the role of E-selectin, Clara cell secretory protein 16 (CC-16), and pulmonary surfactant protein A (SP-A) in the diagnosis of neonatal ARDS.@*Methods@#Full-term newborn with ARDS in 9 hospitals of Jiangsu Province from March 1st 2015 to February 29th 2016 were selected as observation group.According to the lung oxygenation of the neonates, they were divided into three groups: mild, moderate and severe.In addition, 60 normal full-term newborns were selected as control group.In the observation group, venous blood samples were taken on the 1st, 3rd and 7th day of diagnosis and the control group within 7 days after birth.The level of E-selectin, CC-16 and SP-A were detected by double antibody sandwich enzyme-linked immunosorbent assay.The changes of the level of E-selectin, CC-16 and SP-A at different time points and in neonatus with different severity of ARDS were compare with control group and correlatively analyzed.@*Results@#The observation group included 60 newborns who met the diagnostic criteria of ARDS with male 38, female 22, day age (7.3 ±3.3) hours, gestational age (39.5 ±1.7) weeks and birth weight (3280 ±577) g. The control group included 60 normal full-term newborns, with male 30, female 30, day age (6.9 ±4.2) hours, gestational age (39.4 ±1.5) weeks and birth weight (3329 ±593) g. There was no significant difference between two groups.The levels of E-selectin[1 d, 3 d, 7 d: (36.36 ±8.32)μg/L, (45.51 ±9.26)μg/L, (57.15 ±6.84)μg/L], CC-16[1 d, 3 d, 7 d: (25.24 ±8.63)mg/L, (48.33±10.83)mg/L, (18.84±10.11)mg/L]and SP-A [1 d, 3 d, 7 d: (58.38±10.31)mg/L, (53.29±11.31)mg/L, (25.99±6.66)mg/L]in the blood of the observation group increased on the first day and reached the peak on the third day, which were significantly higher than those in the control group [E-selectin, CC-16, SP-A: (15.52 ±6.24)μg/L, (11.26 ±5.18)mg/L, (24.30 ±5.27)mg/L] (P<0.05). The levels of E-selectin [mild, moderate, severe are(30.07±6.10)μg/L, (33.39 ±6.64)μg/L, (41.63 ±7.36)μg/L], CC-16 [mild, moderate, severe are(12.61 ±5.80)mg/L, (25.22 ±6.77)mg/L, (30.61 ±4.69)mg/L]and SP-A [mild, moderate, severe are(49.67 ±8.26)mg/L, (7.11 ±7.94)mg/L, (63.19 ±11.45)mg/L]increased gradually in the blood of ARDS neonates with different severity (P<0.05), especially in moderate and severe degree.There was a significant negative correlation between E-selectin (r=-0.629 8), CC-16 (r=-0.679 3), SP-A (r=-0.458 8) and PaO2/FiO2 (P<0.05).@*Conclusion@#The levels of E-selectin, CC-16 and SP-A in the blood of ARDS neonates increased significantly and were closely related to the severity of the disease, which may be a biomarker of neonatal lung injury.

Progress on PM2.5 and its adverse impact on lung tissues / 中华实用儿科临床杂志

The atmospheric particulate matter(PM) is widely regarded as one of major environmentally and unfriendly ambient air pollution,and therein PM2.5 (diameter≤2.5 μm) is most closely related to human health.Because of its smaller diameter with longer suspension duration,PM can absorb many pathogenic microorganisms,and easily enter into the deep of airway and then deposit on the bronchus and alveoli,and it brings damage to the lung tissues and the surfactant proteins.PM can give rise to oxidative stress,inflammation response,cells and DNA damage.Now,this review focuses on the characterization and composition of PM,as well as the impact of PM2.5 on the lung,surfactant proteins and human health,which helps to call for more people to pay attention to this environmental issue in order to better mitigate and prevent the damage caused by PM.

Protective effects of bone marrow mesenchymal stem cell-derived exosomes on smoke inhalation injury in rats / 医学研究生学报

Objective Acute lung injury induced by variety causes can be reduced by mesenchymal stem cells.Some studies have shown that mesenchymal stem cell-derived exosomes have similar features with mesenchymal stem cell,but its role in acute lung injury is less studied.The study was to investigate the protective role and underlying mechanisms of bone marrow mesenchymal stem cell-derived exosomes (BMSC-DEs) on smoke inhalation injury (SⅡ) in rats.Methods Thirty Wistar rats were randomly divided into 3 equal groups:normal control group,smoke inhalation injury (SⅡ) model group and bone marrow mesenchymal stem cell-derived exosomes (BMSC-DEs) treated group.12 h after establishing the SⅡ model,BMSC-DEs treated group was injected with 0.5 mL BMSC-DEs (derived from 4× 106 BMSCs),and normal control group and SⅡ model group were injected with equivalent volume of normal saline.7 days later,samples were collected.The histopathologic changes of lung were observed after HE staining;BCA was used to test the amounts of total protein in bronchoalveolar lavage fluid (BALF);Enzyme linked immunosorbent assay was used to test the levels of tumor necrosis factor-α (TNF-α) and keratinocyte growth factor (KGF) in the lung tissue;Immunohistochemical was used to test the levels of pulmonary surfactant protein C(SP-C).Results The BALF levels of total protein of SⅡ group was significantly higher than those of normal control group (P＜0.01) and BMSC-DEs groups(P＜0.05);Compared with normal group [(0.164±0.021) ng/L],the levels of tumor necrosis factor-α of SII and BMSC-DEs groups [(0.355±0.106)、(0.234±0.024) ng/L] (P＜ 0.05) were significantly higher,and SⅡ group was higher than that of BMSC-DEs group(P＜0.01);Compared with normal group,the KGF protein expression level in lung tissue of SⅡ group was significantly lower (P＜0.05),but BMSC-DEs group was higher (P＜0.05).BMSC-DEs group was higher than SⅡ group (P＜0.01);Immunohistochemistry showed that the SP-C expression level in lung tissue of SⅡ group was significantly lower than those of other groups (P＜0.05).There was no statistically difference between BMSC-DEs group and control group (P＞0.05).Conclusion BMSC-DEs has a protective effect of smoke inhalation injury rats,the underlying mechanism may be related to BMSC-DEs to reduce inflammation and promote restoration of the alveolar epithelial type Ⅱ.

Determination of Clara cell secretory protein 16 and pulmonary surfactant protein D in children with severe pneumonia and its clinical significance / 临床儿科杂志

Objective To explore the changes of serum Clara cell secretory protein 16 (CC16), pulmonary surfactant protein D (SP-D) in children with pneumonia and its clinical significance. Methods A total of 81 pediatric patients with community-acquired pneumonia were selected, including severe pneumonia with mechanical ventilation group (n=21), severe pneumonia with non-mechanical ventilation group (n=30), mild pneumonia group (n=30), and the control group (n=20) was selected in the physical examination of healthy children over the same period. We detected the concentration of serum CC16, TNF-α, IL-6 and SP-D for the 4 groups by ELISA, and evaluated the clinical values of serum CC16, TNF-α, IL-6 and SP-D for severe pneumonia by using ROC curve.We recorded pulmonary dynamic compliance(Cdyn),airway resistance(Raw),peak inspiratory pressure (PIP), work of breathing (WOB) and other respiratory mechanical parameters, and analyzed the correlations between CC16 and TNF-α, IL-6, SP-D and respiratory mechanical parameters. Results The concentrations of serum CC16 in pneumonia group were all significantly lower than that in the control group, and those in severe pneumonia groups were lower than that in mild pneumonia group, and mechanical ventilation group was lower than that in non-mechanical ventilation; the concentration of serum TNF-α, IL-6 and SP-D in pneumonia groups were all obviously higher than that in the control group, and severe pneumonia group were higher than that in mild pneumonia group, and those in mechanical ventilation group were also higher than that in non-mechanical ventilation group (P<0.05). Compared to that before removing the ventilator, concentration of serum CC16 in severe pneumonia with mechanical ventilation group decreased significantly at 1 hour and lowered down at 72 hours; but the concentration of serum TNF-α, IL-6 and SP-D in severe pneumonia with mechanical ventilation increased significantly at 1 hour and went higher at 72 hours, the differences were all statistically significant (all of P<0.05); compared to that before weaning from the ventilator, the value of Cdyn decreased obviously in severe pneumonia with mechanical ventilation at 72 hours and lowered down at 1 hour; but the values of Raw, PIP, WOB in severe pneumonia with mechanical ventilation increased obviously at 72 hours and more higher at 1 hour, the differences were all statistically significant (all of P<0.05). The concentration of serum CC16 showed all negative correlations with TNF-α, IL-6 and SP-D, but it showed positive correlation with Cdyn(all of P<0.01).In the ROC curve,the area under the ROC curve of CC16,TNF-α,IL-6 and SP-D in serum was 0.905, 0.704, 0.832, 0.825, respectively (for all of which P<0.01). Conclusion The concentrations of serum CC16 and SP-D were associated with the severity of community acquired-pneumonia in children. The level of serum CC16 was positive associated with Cdyn in children with mechanical ventilation. CC16 has better prediction and evaluation effect on the change of severe pneumonia.

Study on the relationship of serum pulmonary surfactant protein and vascular endothelial growth factor related indexes to lung cancer / 国际检验医学杂志

Objective To study the relationship of serum pulmonary surfactant protein and vascular endothelial growth factor re‐lated indexes to lung cancer .Methods Totally 56 patients with lung cancer in our hospital from September 2014 to December 2015 were selected as the observation group ,56 healthy personnel at the same period were selected as the control group ,then the serum pulmonary surfactant protein and vascular endothelial growth factor related indexes of control group and observation group ,obser‐vation group with different stages and pathological classifications were compared ,and the relationship between serum pulmonary surfactant protein and vascular endothelial growth factor related indexes and lung cancer were satisfacted with Logistic analysis .Re‐sults The serum pulmonary surfactant protein and vascular endothelial growth factor related indexes of observation group were all higher than those of control group ,and the serum pulmonary surfactant protein and vascular endothelial grow th factor related inde‐xes of observation group with higher stages were all higher than those of patients with lower stages ,and the levels of patients with adenocarcinoma were all higher than those of patients with squamous cell carcinoma ,and the serum pulmonary surfactant protein and vascular endothelial growth factor related indexes all had close relationship to the lung cancer (all P<0 .05) .Conclusion The serum pulmonary surfactant protein and vascular endothelial growth factor related indexes all have close relationship to the lung cancer ,and those indexes of patients with lung cancer should be paid more attention .

Analysis of pulmonary surfactant protein B exons sequencing in 80 very low birth weight prematures / 中华实用儿科临床杂志

Objective To study the relationship between exon gene polymorphism of pulmonary surfactant pro-tein B(SP -B)and the susceptibility and severity,prognosis of respiratory distress syndrome (RDS).Methods To detect the gene sequence of SP -B exon by adopting the gene sequencing technology,and samples were 80 prematures of very low birth weight in Southern Han Chinese,who were divided into 2 groups,the RDS and the non RDS,and the difference of genotype in SP -B exon in 2 groups was compared.Results There was no significant difference between 2 pretem groups in the aspects of the gestational age,sex,birth weight and delivery mode etc(all P >0.05).Fifty -nine prematures of very low birth weight had exons heterogenesis,and there were 2 types of mutations,V1 :Exon2:c.[5A >C]+[5A >C]or c.[5A >C]+[=];V2:Exon5:c.[428C >T]+[428C >T]or c.[428C >T]+[=].There were 20 cases of type V1 ,1 8 cases of type V2,3 cases of type V1 +V2 in 45 cases of RDS,and there were 1 2 cases of type V1 ,9 cases of type V2,no case of type V1 +V2 in the non RDS group.Comparing the incidence of V1 and V2 in 2 groups,there were all significant differences(χ2 =3.73,5.02;all P 0.05).Conclusion Gene polymorphism of SP -B exon are risk factors for premature of very low birth weight in southern Han Chinese in RDS.

The relationship between vitamin A and pulmonary surfactant protein with neonatal pulmonary function / 中国小儿急救医学

Objective To investigate the correlation between vitamin A and surfactant protein (SP)-B, SP-C in human body,and to explore the effects on lung development and pulmonary function of neonates. Methods We collected the blood samples of 170 pregnant women and umbilical cord serum of their neonatal babies. The levels of vitamin A in pregnant women and their neonatal babies,and the levels of SP-B and SP-C in neonatal umbilical cord serum were detected by ELISA. We conducted a follow-up by standard telephone questionnaire,which we concerned was the number of respiratory tract infection within six months,in order to assess the neonatal pulmonary functions. Results (1) There was a positive correlation between the vitamin A levels in neonatal umbilical cord blood and in the blood of pregnant women(r=0. 866,P<0. 05). (2) There was a positive correlation between the vitamin A levels in neonatal umbilical cord blood and the levels of SP-B,SP-C in the blood(r=0. 817,P<0. 05). (3)In the follow-up of 170 cases of infants within six months,three cases with pneumonia hospitalized more than once,but no respiratory distress syndrome hap-pened. Conclusion Vitamin A can be used as an important biological marker to evaluate the neonatal pul-monary maturity. If we detect the vitamin A levels of pregnant women,increase the intake of vitamin A,we can improve the content of SP-B,SP-C,improve the development of neonatal lung function in growth.

Effect of antioxidant on pulmonary surfactant in acute lung injury rats / 中国小儿急救医学

Objective To explore the changes of neutrophil elastase (NE) and surfactant protein A (SP-A) in acute lung injury(ALI) rats,and the effect of antioxidant. Methods Sixty healthy mature Wister rats were divided into 2 groups, the control group and treatment group. The rats in two groups all received peritoneal injection of E. coli to establish the ALI animal model. 30 minutes after injection of E. coli,the rats in treatment group were injected reduced glutathione from vena caudalis. The levels of NE in blood and expressions of SP-A in lung tissue were detected at 3,6 and 12 hours after injection of E. coli. Results ALI symptom appeared 3 hours after injection of E. coli in the control group, obvious after 6 hours, the rats vomi-ted pink secretion after 12 hours. Lung edema and bleeding were found by pathologic examination. No obvious symptom was found in treatment group after 3 hours, slight tachypnea after 6 hours, slight edema in pulmonary tissue after 12 hours. After administration of reduced glutathione,levels of NE at 3,6 and 12 hours in the treatment group were lower than those in the control group,and indicated statistical significance in 6 and 12 hours(P <0. 05) ;Levels of SP-A in 3,6 and 12 hours in the treatment group were higher than those in the control group, and indicated statistical significance in 3,6 and 12 hours (P < 0. 05). Conclusion Dysfunction of pulmonary surfactant is secondary in ALI, degradation of SP-A is the one of reasons, the application of reduced glutathione as antioxidant, could effectively suppress NE to decompose basosexine elastin of cells and destroy surface active protein, has protective effect on ALI.

The contents of Surfactant Protein A and D in BAL Fluids of Healthy Smokers and Healthy Nonsmokers / 中国基层医药

Objective To observe whether cigarette smoke alter the component of pulmonary surfactant pro-tein A and D. Methods In this study, we determined the contents of SP-A and SP-D in BAL fluids of healthy smok-ers and nonsmokers by enzyme linked immunosorbent assay(ELISA). Resluts The contents of SP-A and SP-D in BAL fluids were significantly decreased in smokers compared to those in nonsmokers[ (3.1±0.40) μg/ml vs (1.8± 0.4)μg/ml, (1.5±0.2) μg/ml vs (0.6±0.1)μg/m, all P <0.05]. Conclusion These results suggested that the decreased levels of SP-A and SP-D in smokers may impair the host defense functions of suffactant in the peripheral airways and might have a crucial role in the development of chronic obstructive lung disease.

Influence of Vascular Endothelial Growth Factor on Expression of Pulmonary Surfactant Protein B in Premature Rats / 实用儿科临床杂志

Objective To study the influence of vascular endothelial growth factor(VEGF) on development of alveolar epithelial cell Ⅱ (AECⅡ) and expression of pulmonary surfactant protein B(SP-B) in premature rats.Methods Wistar rats at 19 days gestation were paunched to get embryo and primary AECⅡculture.The rats were divided into 4 groups ,VEGF-165 group,Dexamethasone group,VEGF and Dexamethason group,control group. AECⅡ and SP-B expression were measured by immunology histochemistry.Results SP-B had positive expression in VEGF group, Dexamethason group, VEGF and Dexamethason group. SP-B had negative expression in control group.Conclusion VEGF-165 can increase SP-B positive expression and secret of AECⅡ.VEGF promotes lung maturity.

Cultivation,identification and sorting of bronchoalveolar stem cells derived from mouse lung / 第三军医大学学报

Objective To cultivate,identify and sort bronchoalveolar stem cells(BASC)derived from normal adult mouse lung.Methods After enzymatic digestion of lung tissue with dispase and collagenase in combination,the Sca-1+ cells were isolated from the obtained pulmonary cells by magnetic cell sorting.These Sca-1+ cells were cultured in dishes coated with collagen and mouse fibroblast cell line Swiss-3T3 under a serum-free culture system for BASC,which were identified by the dual-color immunofluorescent staining clara cell specific antigen(CCA)and surfactant protein C(SP-C).Finally,these pure BASC were isolated by the flow cytometry.Results One lung of normal adult mouse could yield(1.6-1.8)?107 nucleated cells in this enzyme digestion procedure.The percentage of Sca-1+ cells we sorted from lung tissue was much higher than the unsorted [(87.3?5.9)% and(9.6?1.8)%,P

Cloning of SPA gene promoter and analysis of its transcriptional targeting activity / 中国病理生理杂志

AIM:To clone the SPA gene promoter and construct its luciferase report vector of SPA gene and to study its transcriptional targeting activity.METHODS:① The SPA gene sequence was acquired from GenBank,of which the upstream was analyzed according to bioinformatics.The results showed that the upstream region of SPA gene sequence about 163bp has the function of promoter.② The SPA gene promoter fragment was generated by polymerase chain reaction and then subcloned into the multiple clone site(MCS)of luciferase report gene vector pGL3-basic to generate the recombined plasmid pGL3-SPA.This fragment was also subcloned into pGL3-control to generate recombined plasmid pGL3-SPA-enhancer by replacing its primary SV40 promoter.③ pGL3-SPA,pGL3-SPA-enhancer,pGL3-control,pGL3-basic were cotransfected with pRL-TK into A549 cells and H441 cells.The luciferase activities were measured by dual luciferase reportor(DLR)system.RESULTS:Sequencing and restricted digestive results showed that SPA gene promoter was successfully cloned and identified,and also correctly subcloned into plasmid pGL3-basic and pGL3-control to construct its luciferase report plasmid pGL3-SPA and pGL3-SPA-enhancer,respectively.The transcriptional activity was high in H441 cells.CONCLUSION:The luciferase report system of SPA gene promoter is successfully constructed with high transcriptional activity.