The correlation between pulmonary surfactant protein B gene polymorphism and susceptibility to respiratory distress syndrome in Uyghur newborns / 中国新生儿科杂志

Objective:To study the relationship between the 1580 locus (rs1130866) gene polymorphism of pulmonary surfactant protein B (SP-B) gene and susceptibility of neonatal respiratory distress syndrome (RDS) in Uygur newborns.Methods:From June 2019 to May 2020, Uyghur premature infants admitted to the Department of Neonatology of our hospital were prospectively enrolled and assigned into RDS group and control group according to their diagnosis. The genotype and allele distribution of SP-B gene 1580 locus were examined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology. The genotype and allele frequency of SP-B gene 1580 locus in RDS infants among Uyghur, Mongolian and Han nationality were compared.Results:A total of 160 infants were enrolled, including 80 in the RDS group and 80 in the control group. Three genotypes (TT, TC and CC) were detected in both groups. The frequencies of the three genotypes in the RDS group were 18.8%, 53.8% and 27.4%, respectively, with T allele frequency 45.6% and C allele frequency 54.4%. The frequencies of the three genotypes in the control group were 22.5%, 52.5% and 25.0%, respectively, with T allele frequency 48.8% and C allele frequency 51.3%. No significant differences existed in genotype frequency and allele frequency distribution between the two groups ( P>0.05). The CC genotype frequency of SP-B gene 1580 locus in Uyghur was significantly different from Mongolian and Han nationality ( P<0.05). Significant difference existed in C allele frequency between Uyghur and Han nationality ( P<0.05), while no significant difference existed in allele frequency between Uyghur and Mongolian nationality ( P>0.05). Conclusions:No correlation exists between the polymorphism of SP-B gene 1580 locus (rs1130866) and RDS in Uygur premature infants. Different ethnic groups have different SP-B gene 1580 locus polymorphism.

The relationship between bronchopulmonary dysplasia and SP-B intron 4 gene polymorphism in Mongolian and Han newborns in Inner Mongolia / 中国新生儿科杂志

Objective To study the relationship between pulmonary surfactant protein B (SP-B) intron 4 gene polymorphism and bronchopulmonary dysplasia (BPD) in premature infants.Method From January 2016 to January 2019,premature infants diagnosed with BPD in our hospital were selected as the BPD group,and non-BPD premature infants of the same ethnic group were selected as the control group.The genotype and allele distribution of SP-B intron 4 were analyzed using polymerase chain reaction (PCR)method.Result A total of 74 infants with BPD were included,including 30 Mongolian infants and 44 Han infants.A total of 134 cases were in the control group,including 56 Mongolian infants and 78 Han infants.Wild type and variant type (including insertion and deletion) could be detected in SP-B intron 4 gene in both Mongolian and Han infants.The frequencies of wild and variant genotypes and alleles in Mongolian BPD infants were similar with the control group [36.7％ (11/30) vs.19.6％ (11/56),21.7％ (13/60) vs.12.5％ (14/112)] (P ＞ 0.05).The frequencies of wild and variant genotypes and alleles in Han infants with BPD were significantly different from the control group [31.8 ％ (14/44) vs.12.8 ％ (10/78),20.5 ％(18/88)vs.7.1％(11/156)] (P＜0.05).Conclusion The variation of intron 4 gene in SP-B may be related with the genetic susceptibility of Han infants with BPD in Inner Mongolia.

Effects of Mycobacterium tuberculosis heat-resistant antigen on secretion of SP-B and apoptosis in A549 cells / 安徽医科大学学报

Objective To investigate secreting pulmonary surfactant-associated protein B (SP-B) and apoptosis in human lung adenocarcinoma cell line (A549) cells,treated with heat-resistant antigen of Mycobacterium tuberculosis (Mtb-HAg).Methods Different concentrations of Mtb-HAg were used to culture A549 cells for 24 h and 48 h respectively,and the blank control groups(control group 1 and control group 2) and positive control groups [lipopolysaccharide (LPS) group and curcumin group] were set up.SP-B in the culture supernatant,was assessed by ELISA in control group 1,LPS group and experimental group 1 (A549 cells were respectively induced by 2 μg/ml,3 μg/ml Mtb-HAg to culture for 24 h and 48 h).For control group 1,LPS group and experimental group 1,the relative expression of SFTPB gene was quantified with quantitative real-time fluorescence quantitative PCR (qRT-PCR).The apoptosis rate of control group 2,curcumin group and experimental group 2(A549 cells were respectively induced by 2 μg/ml,3 μg/ml Mtb-HAg to culture for 24 h) in A549 cells were detected by flow cytometry.Results SP-B expression in the experimental group 1 was significantly lower than the control group 1,the difference was statistically significant (P ＜ 0.05).The difference of SP-B expression in the experimental group 1 was not obvious with the prolongation of the same concentration.At the same incubation time,the expression of SFTPB in the experimental group 1 decreased obviously with the increasing concentration of Mtb-HAg,the difference was statistically significant (P ＜ 0.05).The change with time was not significant.The apoptosis rate of curcumin group and experimental group 2 were significantly higher than that in control group 2 in A549 cells (P ＜ 0.05).Conclusion Mtb-HAg inhibited the expression of SP-B in A549 cells significantly,and induced apoptosis of A549 cells.

Fetal lung structures and expression of pulmonary surfactant proteins of B and C and their modulating factors in rat with gestational diabetes mellitus / 中华围产医学杂志

Objective To explore the lung developmental disorder of rats with gestational diabetes mellitus (GDM) via investigating the GDM rat fetal lung structures and expression of pulmonary surfactant proteins (SP)-B,SP-C,thyroid transcription factor (TTF)-1 and pleiomorphic adenoma gene like (PLAGL)-2.Methods Sprague-Dawley rats were used to construct the GDM model.Twenty GDM rats were used as GDM group and 20 normal pregnant rats as control group.Cesarean section was performed on day 21 of gestation and random blood sugar was detected,and fetal rats were counted and weighed.Ultrastructure of the fetal lungs was studied by transmission electron microscopy.Sixty fetal rats were selected randomly in each group,and 360 paraffin sections were made from fetal lungs.One hundred discontinuous paraffin sections were picked up in each group to observe morphological and structural changes under optical microscope.The other one hundred discontinuous paraffin sections were picked up in each group to detect the location and expression of SP-B,SP-C,TTF-1 and PLAGL-2 protein by immunohistochemistry.Nine fetal rats were selected randomly to detect the expression level of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in fetal lung tissues by Western blotting.Twenty seven fetal rats were selected randomly to detect the mRNAs level of SP-B,SP-C,TTF-1 and PLAGL-2 by real-time quantitative polymerase chain reaction.Independent sample t-test was used for statistical analysis.Results The average random blood glucose level in GDM group was significantly higher than that in control group [(26.8± 2.8) vs (4.9± 0.5) mmol/L,t=-34.05,P=0.00].The average weight of fetal rats in GDM group was higher than that in control group [(5.6±0.6) vs (5.2±0.5) g,t=-1.97,P=0.03].Alveolar number (10.1 ± 1.6 vs 12.1 ± 1.3) and alveolar area [(986.9 ± 5.5) vs (1 257.3± 5.0) μ m2] in GDM group was less than that in control group (t=9.84 and 27.53,both P ＜ 0.05).Alveolar septum [(11.5±6.2) vs (9.9±4.3) μm] in GDM group was higher than that in control group (t=-2.17,P ＜ 0.05).Microvillus in type] cells were short and the number of lamellar bodies was significantly decreased in GDM group.SP-B,SP-C,TTF-1 and PLAGL-2 proteins were distributed in the cytoplasm in granular form.The average value of absorbance of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in GDM group was 1.15±0.12,1.23±0.06,0.87±0.21 and 1.21 ±0.18 respectively;and that in control group was 1.22±0.05,1.31 ±0.14,1.12±0.09 and 1.33 ±0.07 respectively.The value in GDM group was lower than that in control group (t=2.40,2.35,4.89,and 2.77 respectively,all P ＜ 0.01).The expression level of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in GDM group was 0.57± 0.09,0.45±0.03,1.50±0.04 and 1.11 ±0.04 respectively;and that in control group was 0.81 ±0.03,0.66±0.04,1.69±0.05 and 1.46±0.07 respectively.The value in GDM group was lower than that in control group (t=1 1.77,11.09,8.80 and 13.37,respectively,all P ＜ 0.01).The mRNA level of SP-B,SP-C,TTF-1 and PLAGL-2 in GDM group was 0.60±0.04,0.79±0.04,0.81 ±0.03 and 0.79±0.05 respectively;and that in control group was 1.06±0.19,1.03±0.24,1.03±0.18 and 1.02±0.19 respectively.The value in GDM group was lower than that in control group (t=6.80,2.98,3.54 and 3.54 respectively,all P ＜ 0.01).Conclusions The protein expression level of SP-B and SP-C in fetal lungs of GDM rats decreases obviously,possibly because of the down-regulation of the gene expression of TTF-1 and/or PLAGL-2.The pathological changes in fetal lungs of GDM rats might be associated with the descending level of SP-B and SP-C protein.

High postoperative serum levels of surfactant type B as novel prognostic markers for congenital heart surgery / Níveis séricos pós-operatórios tipo surfactante B elevados como novos marcadores prognósticos para cirurgia cardíaca congênita

Objective: Congenital heart diseases are observed in 5 to 8 of every 1000 live births. The presence of a valuable biomarker during the surgical periods may aid the clinician in a more accurate prognosis during treatment. Methods: For this reason, surfactant protein B plasma levels may help to evaluate patients with cardiac problems diminishing the alveolocapillary membrane stability. In this study, plasma levels of this biomarker were measured in the preoperative and postoperative periods. This study was conducted to detect the differences between pulmonary hypertensive and normotensive patients. The differences before and after cardiopulmonary bypass were examined. Results: The differences in cardiopulmonary bypass time, cross-clamp time , inotropic support dose, and duration of intensive care of patients with and without pulmonary hypertensive were found to be statistically significant (P<0.05). The results revealed that this pathophysiological state was related to other variables that were studied. We believe that the differences in preoperative and postoperative SPB levels could be attributed to alveolocapillary membrane damage and alveolar surfactant dysfunction. We found that this pathophysiological condition was significantly associated with postoperative parameters. Conclusion: The findings of the current study showed that surfactant protein B was present in the blood of patients with a congenital heart disease during the preoperative period. Long by-pass times may exert damage to the alveolocapillary membrane in patients with pulmonary hypertension and preoperative heart failure, and it is recommended to keep the option of surfactant therapy in mind during the postoperative course at the intensive care unit before preparing the patients for extubation. .

Objetivo: As cardiopatias congênitas são observadas em 5 a 8 em cada 1.000 nascidos vivos. A presença de um biomarcador importante durante os períodos cirúrgicos pode auxiliar o clínico a um prognóstico mais preciso durante o tratamento. Métodos: Por esta razão, os níveis plasmáticos de proteína B do surfactante podem ajudar a avaliar os pacientes com problemas cardíacos, diminuindo a estabilidade da membrana alvéolo-capilar. Neste estudo, os níveis plasmáticos deste biomarcador foram medidos nos períodos pré-operatório e pós-operatório. Este estudo foi realizado para detectar as diferenças entre pacientes hipertensos e normotensos em nível pulmonar. As diferenças antes e depois da circulação extracorpórea foram examinadas. Resultados: As diferenças no tempo de circulação extracorpórea, tempo de pinçamento, a dose de drogas vasoativas, e a duração da terapia intensiva de pacientes com e sem hipertensão pulmonar foram estatisticamente significativas (P<0,05). Os resultados revelaram que este estado fisiopatológico foi relacionado a outras variáveis que foram estudadas. Acreditamos que as diferenças nos níveis de SPB pré-operatório e pós-operatório pode ser atribuída a danos na membrana alvéolo-capilar e disfunção do surfactante alveolar. Descobrimos que esta condição fisiopatológica foi significativamente associada com parâmetros pós-operatórios. Conclusão: Os resultados do estudo mostraram que a proteína B surfactante estava presente no sangue de pacientes com doença cardíaca congênita no pré-operatório. Longos tempos de circulação extracorpórea podem exercer danos na membrana alvéolo-capilar em pacientes com ...

Effects of antenatal dexamethasone and ambroxol on expression of surfactant protein-B mRNA and ;thyroid transcription factor-1 in pulmonary tissue of premature rats / 中华围产医学杂志

To explore the effects and mechanism of dexamethasone and ambroxol on expression of surfactant protein (SP)-B mRNA and thyroid transcription factor (TTF)-1 in premature rat lung. Methods Sixteen pregnant Sprague-Dawley rats were randomly divided into four equal groups: two doses of dexamethasone (0.2 mg/kg injected intramuscularly on Day 17 and 18 of pregnancy respectively);single dose of dexamethasone (0.2 mg/kg injected intramuscularly on Day 18 of pregnancy);ambroxol group (100 mg/kg injected intraperitoneally on Day 16, 17 and 18 of pregnancy respectively); and control group (normal saline injected intraperitoneally on Day 16, 17 and 18 of pregnancy respectively). There were four pregnant rats in each group. All of the fetal rats were taken out on Day 19 of pregnancy as the preterm birth model, and 20 fetal rats from each group were randomly selected. The ratio of body weight to fetal lung weight of newborn rats was calculated. Changes in lung morphology were observed under light microscopy and the ratio of alveoli surface area to alveolar septae surface area was calculated. Expression of TTF-1 protein was determined by immunohistochemistry. Expression of SP-B mRNA was detected by reverse transcriptase-polymerase chain reaction. One-way analysis of variance, Student-Newman-Keuls method and Pearson correlation analysis were applied as statistical methods. Results (1) The ratio of body weight to fetal lung weight was (6.5±0.6), (7.9±0.8), (9.5±0.8) and (9.5±0.9) mg/g in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=67.69,P<0.01). The ratio of two doses and one dose of dexamethasone group was lower than that of control group (q=17.143 and 9.143, all P<0.01) and ambroxol group (q=17.143 and 9.143, all P<0.01). The ratio of two doses dexamethasone group was lower than that of one dose dexamethasone group (q=8.000, P<0.01). (2) The ratio of alveoli surface area to alveolar septae surface area was 2.19±0.15, 1.70±0.18, 1.67±0.13 and 1.08±0.12 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=190.85, P<0.01). The ratio of two doses of dexamethasone group, one dose of dexamethasone group and ambroxol group were higher than that of the control group (q=33.639, 18.788 and 17.879, all P<0.01). The ratio of two doses dexamethasone group was higher than that of one dose dexamethasone group (q=14.848, P<0.01). (3) Expression of TTF-1 protein was 0.311±0.018, 0.224±0.019, 0.196±0.013 and 0.191±0.018 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=211.69,P<0.01). TTF-1 protein expression of two doses and one dose of dexamethasone group were higher than that of control group (q=30.000 and 8.250, all P<0.01) and ambroxol group (q=28.750 and 7.000, all P<0.01). TTF-1 protein expression of two doses dexamethasone group was higher than that of one dose dexamethasone group (q=21.750, P<0.01). (4) Expression of SP-B mRNA was 1.25±0.13, 1.15±0.12, 1.10±0.10 and 1.01±0.12 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=14.48, P<0.01). SP-B mRNA expression of two doses of dexamethasone group, one dose of dexamethasone group and ambroxol group were higher than that of control group (q=9.231, 5.385 and 3.462, all P<0.01). SP-B mRNA expression of two doses of dexamethasone group was higher than that of ambroxol group (q=5.769, P<0.01) and one dose of dexamethasone group (q=3.846, P<0.01). (5)TTF-1 expression in two doses of dexamethasone group, one dose of dexamethasone group and control groups was positively correlated with SP-B mRNA expression (r=0.512, 0.597 and 0.449, respectively, all P<0.05). Conclusions Ambroxol can accelerate the maturation of fetal lung with minimal adverse effects on fetal lung weight. Ambroxol might be an alternative to dexamethasone to prevent neonatal respiratory distress syndrome.

Construction of eukaryotic cell expression vectors pIRES2-EGFP-SP-B-C/T 1580 and evaluation of their expressions in 293T cells / 中国医师杂志

Objective To construct two kinds of eukaryotic ccll expression vcctors pIRES2-EGFP-SP-B-C/T 1580 and evaluate their expressions in 293T cells,for the further study of relationship between polymorphism of surfactant protein B (SP-B) gene and bronchopulmonary dysplasia (BPD).Methods The eukaryotic pIRES2-EGFP-SP-B-C/T 1580 expression vectors were constructed by gene recombination,and identified by gene sequencing.The recombinant expression vectors were transfected into 293T cells by lipofectamine2000.The expression of green fluorescence protein in 293T cells was observed by fluorescence microscopy.The mRNAs and proteins of SP-B-C/T 1580 were tested and identified by reverse transcriptionpolymerase chain reaction-restriction fragment length polymorphism(RT-PCR-RFLP) and western blot.Results Two recombinant plasmids contained the complete cDNA of SP-B with the same sequence as in gene bank.The base of SP-B 1580 gene of pIRES2-EGFP-SP-B-C 1580 was C,that of pIRES2-EGFP-SP-B-T 1580 was T.After being transfected into 293T cells,highly efficient expression of SP-B-C/T 1580 gene was detected at mRNA and protein levels.Conclusions The pIRES2-EGFP-SP-B-C/T 1580 eukaryotic cell expression vectors were successfully constructed.

Gene polymorphism in intron 4 of surfactant protein-B in bronchopulmonary dysplasia infants / 中华围产医学杂志

ObjectiveTo investigate the change of gene polymorphorism of surfactant protein-B (SP-B) intron 4 in infants with bronchopulmonary dysplasia (BPD).MethodsForty-five infants with BPD (BPD group) and ninety-nine infants without lung diseases (control group) who admitted into Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology from July 2008 to July 2011 were selected into this study.Genotyping for fragment length polymorphism of SP-B intron 4 was performed by polymerase chain reaction (PCR),agarose gel electrophoresis,cloning and sequencing methods in both groups.Differences of allele frequencies (invariant allele and variant allele) and genotype frequencies (invariant genotype and variant genotype) between BPD group and control group were analyzed.The differences of gestational age and birth weight between the two groups were compared with Independent-Samples t test.The gender composition and differences of allele or genotype frequencies between the two groups were compared with Chi-square test.Results Invariant allele frequencies in BPD group and control group were 83.3％ (75/90) and 92.0％ (182/198),and variant allele frequencies were 16.7％ (15/90,including eight insertion alleles and seven deletion alleles) and 8.1％ (16/198,including eight insertion alleles and eight deletion alleles).There were significant differences between the two groups (x2 =4.75,P =0.029).In BPD group,there were 32 cases (71.1 ％,32/45) invariant genotypes and 13 cases (28.9 ％,13/45,including seven cases insertions and six cases deletions) variant genotypes; in the control group,there were 85 cases invariant genotypes (85.8％,85/99) and 14 cases (14.1％,14/99,six insertions and eight deletions) variant genotypes.Significant difference was found between the two groups (x2=4.42,P＜0.036). ConclusionsVariations of SP-B intron 4 were more in BPD infants,and the variation of SP-B intron 4 might be associated with BPD.

Roles of vascular endothelial growth factor and dexamethasone in expressions of surfactant protein B and transforming growth factor-β1 in type Ⅱ alveolar epithelial cells / 中华围产医学杂志

ObjectiveTo investigate the effects of vascular endothelial growth factor (VEGF) and dexamethasone on mRNA expressions of surfactant protein B (SP-B) and transforming growth factor-β1 (TGF-β1) of type Ⅱ alveolar epithelial cell (AECⅡ). MethodsAECⅡ were isolated and purified from fetal rat lung tissues,then cultured with different dose of VEGF (25,50 and 100 ng/ml) and dexamethasone (25,50,100 and 200 nmol/ml).The mRNA levels of SP-B and TGF-β1 were detected by real-time quantitative polymerase chain reaction (RT-PCR) and expression of TGF-β1 protein was detected by immunocytochemistry.ANOVAor q-test wasappliedtocompare the difference among groups.ResultsCompared with control group,SP-B mRNA levels in 25 ng/ml VEGF group and 25,50,100 and 200 nmol/ml dexamethasone groups were higher (13.500±3.172,3.547±0.690,5.219±0.782,3.493±0.335,and 3.981 ± 1.133 vs 1.001 ± 0.059,q=-5.286,-4.943,- 7.228,- 9.906 and - 3.525 respectively,P＜0.05) ; TGF-β1 mRNA expression of 25 ng/ml VEGF group,50,100 and 200 nmol/ml dexamethasone group was lower (0.451 ± 0.078,0.579±0.019,0.422 ± 0.020 and 0.769 ± 0.025 vs 1.019±0.226,q=4.110,3.356,4.551 and 1.901 respectively,P＜0.05) ; other groups had no significant differences compared with control group (P＞0.05).Immunocytochemistry showed that the positive rate of TGF-β1 expression in 25 ng/ml VEGF,50,100 and 200 nmol/ml dexamethasone group was 23％,26％,22％ and 29％,respectively,while in the control group,the expression of TGF-β1 was positive in most of the AECⅡ (80％).ConclusionsBoth VEGF and dexamethasone could increase the expression of SP-B at mRNA level at appropriate concentrations.At the same time,the expression of TGF-β1 is inhibited.It is suggested that both VEGF and dexamethasone might increase the mRNA expression of SP-B by inhibiting the expression of TGF-β1.

Effects of different ventilation modes on efficacy of exogenous pulmonary surfactant for treatment of rats with ventilator-induced lung injury / 中华麻醉学杂志

Objectlve To investigate the effects of different ventilation modes on the efficacy of exogenous pulmonary surfactant(PS)for the treatment of rats with ventilator-induced lung injury(VILI).Methods Forty-two male Wistar rats weighing 310-356 g were randomly divided into 6 groups(n=7 each):group CVT6,group SVT6,group CVT10,group SVT10,group CVT14 and group SVT14.The tidal volume(VT)was set at 6,10 and 14 ml/kg respectively and the respiratory rate(RR) was 75,45 and 32 bpm respectively.The animals were anesthetized with intraperitoneal 3% Pentobarbital 50 mg/kg,then tracheostomized and intubated.VILI model was induced by high-pressure ventilation (HPV) with peak inspimtory pressure (PIP) 40 cm H2O and without positive end-expiratory pressure (PEEP).The air was injected into the trachea via the airway at the end ofexpiration before HPV (T0,baseline value) and 15-25 min of HPV,the airway pressure monitored and the lung compliance(C) calculated.When C was decreased to half of the baseline value,PEEP was increased to 7.5 cm H20.After the tracheal edema fluid was removed,the PS 100 mg/kg was immediately injected into the trachea in group SVT6,SVT10 and SVT14.The equal volume of air was injected into the trachea in group CVT6,CVT10 and CVr14 instead of PS.Then the rats in different groups were ventilated with the corresponding ventilation modes.MAP was monitored and blood samples were token from femoral artery for blood gas analysis at T0, 5 min after HPV (T1 ), and 15, 30, 60, 90, 120 min (T2-6) after administration of PS. The tracheal edema fluid was collected at T1 and T6.The rats were killed at T6 and the lung tissues taken for microscopic examination. Results With the same ventilation mode, the VILI was significantly alleviated after administration of PS. With different ventilation modes,the lung injury was significantly reduced in group SVT 10 compared with the other groups. Conclusion The efficacy of PS for the treatment of rats with VILI is good using the ventilation strategy with VT of 10 ml/kg and RR of 45 bpm.