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Objective:To evaluate the role of epidermal growth factor (EGF) in repair of lung tissues in mice with acute respiratory distress syndrome (ARDS).Methods:Fifty SPF male C57BL/6 mice, aged 6-8 weeks, weighing 21-23 g, were divided into 5 groups ( n=10 each) using a random number table method: control group (group C), EGF group, LPS+ PBS group, LPS+ EGF group and AG1478+ LPS+ EGF group.PBS 0.1 ml was intraperitoneally injected in group C. EGF 10 μg (0.1 ml) was intraperitoneally injected in group EGF.The equal volume of PBS and EGF 10 μg was intraperitoneally injected at 12 h after tracheal infusion of LPS in group LPS+ PBS and group LPS+ EGF, respectively.EGF receptor (EGFR) antagonist AG1478 1 mg was intraperitoneally injected, 30 min later LPS was tracheally instilled, and 12 h later EGF 10 μg was intraperitoneally injected in group AG1478+ LPS+ EGF.ARDS model was developed by endotracheal instillation of LPS 3 mg/kg.The mice were sacrificed on the 1st and 5th days after development of the model, and lung tissues were obtained for microscopic examination of the pathological changes which were scored after HE staining.Bronchoalveolar lavage was performed on 5th day after development of the model and before sacrifice, and bronchoalveolar lavage fluid (BALF) was collected to detect total protein concentration (by BCA method) and IL-6 and TNF-α concentrations (by enzyme-linked immunosorbent assay). Lung tissues were obtained for determination of the wet/dry lung weight ratio (W/D ratio), expression of lung surfactant associated protein C (SP-C) and proliferating nuclear antigen (PCNA) (by immunofluorescence method), and expression of EGFR, phosphorylated EGFR (p-EGFR), protein kinase B (Akt), and phosphorylated Akt (p-Akt) (by Western blot). Results:Compared with group C, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly increased, the number of cells co-expressing SP-C and PCNA was increased, and p-EGFR/EGFR and p-Akt/Akt ratios were increased in group LPS+ PBS ( P<0.01), and no significant change was found in the indexes mentioned above in group EGF ( P>0.05). Compared with group LPS+ PBS, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly decreased, the number of cells co-expressing SP-C and PCNA was increased, and p-EGFR/EGFR and p-Akt/Akt ratios were increased in group LPS+ EGF ( P<0.01). Compared with group LPS+ EGF, the pathological score, W/D ratio, concentrations of total protein, IL-6 and TNF-α in BALF and neutrophil count were significantly increased, the number of cells co-expressing SP-C and PCNA was decreased, and p-EGFR/EGFR and p-Akt/Akt ratios were decreased in group AG1478+ LPS+ EGF ( P<0.01). Conclusions:EGF can promote the repair of lung tissues in mice with ARDS, and the mechanism may be related to activation of EGFR signaling pathway and promotion of proliferation of alveolar epithelial cell type Ⅱ.
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Objective:To investigate the effects of hypertensive disorders during pregnancy (HDP) on preterm newborns in terms of umbilical cord blood serum ferritin (SF), hemoglobin (Hb) at birth and outcome.Methods:Among inpatients of the First Maternity and Infant Health Hospital Affiliated to Tongji University from October 1, 2015 to December 31, 2016, totally 1 419 cases of preterm newborns were prospectively collected.Preterm infants whose mothers with diagnosis of HDP were recruited as the HDP group.Meanwhile, premature newborns whose mothers without HDP were recruited as the control group.Umbilical cord blood SF levels, Hb levels at birth, outcome of preterm newborns and the basic information for maternity were compared between the two groups.The data of normal distribution between the two groups were compared by independent sample t test.The count data was tested by χ2, and the count data with frequency <5 was tested by Fisher′ s exact test. Results:SF levels of HDP group were significantly lower than the control group [(85.6±67.2) μg/L vs. (103.9±95.5) μg/L]. But Hb levels of HDP group were much higher than the control group [(206.2±33.8) g/L vs. (193.2±31.9) g/L]. The difference between two groups was statistically significant ( t=2.791, 4.825 all P<0.05). Umbilical cord blood SF levels were negatively correlated with Hb levels at birth ( r=-0.120, P<0.001). Moreover, compared to the control group, statistically significant lower incidence of neonatal respiratory distress syndrome (NRDS), pneumonia and bronchopulmonary dysplasia (BPD) in HDP group was observed (all P<0.05). Conclusions:HDP was correlated with umbilical cord blood SF levels and Hb levels at birth in premature newborns.Higher Hb levels and relatively lower incidences of NRDS, pneumonia and BPD were observed in these newborns delivered by mothers with diagnosis of HDP.
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Anemia is one of the common complications in preterm infants.Although there are many factors that can cause anemia of prematurity (AOP), erythropoietin (EPO) deficiency is one of the main causes of AOP.Insufficient EPO levels in premature infants and their mechanisms are the hotspots of AOP research at home and abroad.The rational use of EPO for prevention and treatment of AOP has become an international consensus and significant clinical outcomes have been obtained.The recent progress in the field of AOP, the relationship between EPO and AOP, and the clinical application and efficacy of EPO in the prevention and treatment of AOP in recent 3 to 5 years have been reviewed.
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Anemia is one of the common complications in preterm infants.Although there are many factors that can cause anemia of prematurity (AOP),erythropoietin (EPO) deficiency is one of the main causes of AOP.Insufficient EPO levels in premature infants and their mechanisms are the hotspots of AOP research at home and abroad.The rational use of EPO for prevention and treatment of AOP has become an international consensus and significant clinical outcomes have been obtained.The recent progress in the field of AOP,the relationship between EPO and AOP,and the clinical application and efficacy of EPO in the prevention and treatment of AOP in recent 3 to 5 years have been reviewed.
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Objectives To observe the effect of amphiregulin (Areg) via lateral ventricle injection on focal cerebral ischemia/reperfusion (I/R) injury in rats and to investigate its possible mechanism. Methods A total of 96 3-month old health specified pathogen free SD rats were randomly divided into 6 groups (n=16 in each group):sham operation group (sham group),only exposure of common carotid artery and bifurcation;I/R group,making I/R model;solvent control group,lateral ventricle injection of standard protein solution(5 μl);Areg group,lateral ventricle injection of Areg(2 μg/5 μl);AG1478 group [AG1478,a blocker of Areg receptor epidermal growth factor receptor(EGFR),lateral ventricle injection of AG1478 (2.5 μg/5 μl);Areg combined AG1478 (AAG) group,lateral ventricle giving AG1478 (2.5 μg/5 μl),and then giving Areg (2 μg/5 μl) after 30 mm.The model of focal cerebral I/R injury was induced after 30 min administration of the above last 4 groups.After 24 h of reperfusion,the volume of cerebral infarction, the neurobehavioral score and the number of apoptotic cells in the brain tissue were compared among the groups. After 6 h of reperfusion,the phosphorylation levels of EGFR and protein kinase B(Akt)in ischemic brain tissue were detected. Results Compared with the sham group,the cerebral infarction volume and the number of apoptotic cells in brain tissue were increased significantly,while the neurobehavioral score was decreased(all P<0.05).Compared with the I/R group,the volume of cerebral infarction,the number of apoptotic cells in the brain tissue were decreased significantly,and the neurobehavioral score was increased in the Areg group,the levels of EGFR and Akt phosphorylation were significantly higher (all P <0. 05). Compared with the I/R group,the volume of cerebral infarction and the number of apoptotic cells of the AG1478 group were increased,the levels of EGFR and Akt phosphorylation were decreased(all P<0.05);Compared with the Areg group,the volume of cerebral infarction and the number of apoptotic cells of the AAG group and AG1478 group were increased significantly,and the levels of EGFR and Akt phosphorylation were decreased significantly(all P<0.05). Conclusions Areg reduces the infarct volume in ischemic brain tissue,improves nerve function,and inhibits apoptosis by activating EGFR-Akt signaling pathway. Therefore,it has some protective effect for cerebral I/R injury.
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AIM:To investigate the expression of visfatin in the placenta of patients with preeclampsia and its significance.METHODS:The pregnant women (n=100) were divided into normal pregnancy group , mild preeclampsia group and severe preeclampsia group according to the severity of the disease .The pathological changes of the placenta were observed by hematoxylin-eosin staining .The expression of visfatin at mRNA and protein levels in the placenta was detected by real-time PCR and immunohistochemistry respectively .RESULTS:Compared with normal pregnancy group , the patho-logical changes of the placenta in preeclampsia groups was significant , showing that the structure and the form were disor-dered and incompleted in both cytotrophoblasts and syncytiotrophoblasts .The proliferation of cytotrophoblasts and the num-bers of the placental villi with syncytial knots were observed .The vascular numbers of villi were decreased and congested . The results of immunohistochemistry and real-time PCR showed that the visfatin protein was observed in the cytoplasm of cy-totrophoblasts and syncytiotrophoblasts among 3 groups.The expression of visfatin at mRNA and protein levels was in-creased with the severity of the preeclampsia .CONCLUSION:The injury and dysfunction of vascular endothelial cells in the villi exist in the patients with preeclampsia .High expression of visfatin at mRNA and protein levels in placenta of the patients with preeclampsia indicates that visfatin is closely related to preeclampsia .
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AIM:To investigate the effect of cyclopamine on Hedgehog (HH) signaling, phenotypic transfor-mation and matrix accumulation induced by aristolochic acid (AA) in renal tubular epithelial cell NRK-52E.METHODS:NRK-52E cells were randomly divided into control group (treated with solvent only), AA group (treated with AA at con-centrations of 1, 5, 10 mg/L) and cyclopamine group (treated with AA at concentration of 10 mg/L plus cyclopamine at concentrations of 1, 5, 10μmol/L).After cultured for 24 h, the mRNA expression of Ptch1, Smo,α-SMA, E-cadherin, ZO-1, BMP-7, type I collagen and type III collagen was quantified by real-time PCR.The protein levels of Shh and TGF-β1 were detected by ELISA .Immunofluorescence staining was used to evaluate the expression of Ptch 1, Smo,α-SMA, E-cadherin and type III collagen in the NRK-52E cells.RESULTS: AA increased the expression of TGF-β1, α-SMA and type III collagen, decreased the expression of E-cadherin and ZO-1 protein, and down-regulated the expression of Ptch1, Shh and Smo mRNA in the NRK-52E cells, indicating that AA activated HH signaling , and phenotypic transformation and matrix accumulation occurred in AA-treated NRK-52E cells.Treatment with cyclopamine inhibited HH signaling by decrea-sing Smo expression and increasing Ptch 1 expression.Moreover, cyclopamine also down-regulated the expression of TGF-β1,α-SMA, type I collagen and III collagen , and up-regulated the expression of BMP-7, ZO-1 and E-cadherin.CON-CLUSION:AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells , which can be inhibited by cyclopamine treatment .The possible mechanism is that cyclopamine suppresses the activation of HH signaling , resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition .
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Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.
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Objective: To revise the Online Game Addiction Inventory(OGAI) and examine its psychometric properties.To explore the relationship between OGAI and Self Efficacy of middle school students.Methods: A sample of 1299 middle school students completed the OGAI and Self Efficacy Inventory(SEI).Results: With exploring factor analysis,4 factors were extracted,i.e.Addictive Behavior,Emotional Arousal,Shame & Dissatisfaction and Dysfunction.The reliability of the revised inventory was 0.93 and that of 4 sub-scales ranged from 0.70 to 0.92.Addictive Behavior,Shame & Dissatisfaction and Dysfunction were negatively correlated with Self Efficacy.Emotional Arousal did not correlate with OGAI.Conclusion: The revised Online Game Addiction Inventory(OGAI) is a reliable and valid measurement.OGAI and SEI are significantly related.