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Objective:To explore the value of body temperature, pediatric clinical illness score(PCIS), white blood cell count (WBC), plasma C-reactive protein (CRP), procalcitonin (PCT) and pro-adrenomedullin (pro-ADM) in predicting nosocomial infection in PICU.Methods:From June 2016 to March 2017, the critically ill children in PICU of Children′s Hospital of Fudan University were selected and divided into nosocomial infection group and non nosocomial infection group according to the diagnostic criteria of nosocomial infection.The body temperature, PCIS, WBC, CRP, PCT and pro-ADM were recorded at 4 hours (T1), (48±1) hours (T2), (120±1) hours (T3) and (192±1) hours (T4) after admission, and their predictive value of each index, which was the closest time point (Th) to nosocomial infection was analyzed.Receiver-operating characteristic (ROC) curves were performed to calculate the areas under the curves (AUC), sensitivity and specificity, and multivariate Logistic regression analysis was used to study the risk factors of nosocomial infection.Results:A total of 85 cases were included, including 27 cases in nosocomial infection group and 58 cases in non nosocomial infection group.There was no significant difference in age, weight, body temperature, WBC, PCT, pro-ADM, primary disease and invasive operation between two groups (all P>0.05). There were significant differences in gender, PCIS, CRP, intubation rate and central venous catheterization rate ( P<0.05), when patients were admitted to PICU.At Th, the differences of body temperature, PCIS, CRP, PCT and pro-ADM between two groups were statistically significant ( P<0.05), as well as the AUC were 0.787, 0.755, 0.709, 0.704 and 0.809, respectively, as well as the best cut-off values for predicting nosocomial infection were 38.0 ℃, 87 points, 14.5 mg/L, 0.28 ng/mL and 0.67 nmol/L, respectively.There was no significant difference regarding WBC between two groups ( P>0.05). PCIS may be an independent risk factor for nosocomial infection( OR=0.978, 95% CI 95.9-99.9, P<0.05). Conclusion:Pro-ADM has high sensitivity and specificity in predicting nosocomial infection, and PCIS is an independent risk factor for nosocomial infection.
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OBJECTIVES@#To study the role of adrenomedullin (ADM) in hyperoxia-induced lung injury by examining the effect of ADM on the expression of calcitonin receptor-like receptor (CRLR), receptor activity-modifying protein 2 (RAMP2), extracellular signal-regulated kinase (ERK), and protein kinase B (PKB) in human pulmonary microvascular endothelial cells (HPMECs) under different experimental conditions.@*METHODS@#HPMECs were randomly divided into an air group and a hyperoxia group (@*RESULTS@#Compared with the air group, the hyperoxia group had significant increases in the mRNA and protein expression levels of ADM, CRLR, RAMP2, ERK1/2, and PKB (@*CONCLUSIONS@#ERK1/2 and PKB may be the downstream targets of the ADM signaling pathway. ADM mediates the ERK/PKB signaling pathway by regulating CRLR/RAMP2 and participates in the protection of hyperoxia-induced lung injury.
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Humans , Adrenomedullin/genetics , Endothelial Cells , Hyperoxia/complications , Lung Injury , Receptor Activity-Modifying ProteinsABSTRACT
Abstract Objective: We aimed to investigate the protective effect of adrenomedullin (ADM) on cerebral tissue of rats with cerebral ischemia/reperfusion (I/R) injury. Methods: Thirty-two Wistar rats were randomized into four groups (n=8). In the I/R Group, bilateral common carotid arteries were clamped for 30 minutes and, subsequently, reperfused for 120 minutes. In the ADM Group, rats received 12 µg/kg of ADM. In the I/R+ADM Group, bilateral common carotid arteries were clamped for 30 minutes and, subsequently, the rats received 12 µg/ kg of ADM. Then, reperfusion was performed for 120 minutes. The Control Group underwent no procedure. Blood and brain tissue samples were collected for biochemical and histopathological analysis. Serum malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were analysed. Brain tissue was evaluated histopathologically and neuronal cells were counted in five different fields, at a magnification of ×400. Results: Brain MDA in I/R Group was significantly higher than in ADM Group. Brain GPx and SOD in I/R+ADM Group were significantly higher than in I/R Group. The number of neurons was decreased in I/R Group compared to the Control Group. The number of neurons in I/R+ADM Group was significantly higher than in I/R Group, and lower than in Control Group. Apoptotic changes decreased significantly in I/R+ADM Group and the cell structure was similar in morphology compared to the Control Group. Conclusion: We demonstrated the cerebral protective effect of ADM in the rat model of cerebral I/R injury after bilateral carotid artery occlusion.
Subject(s)
Animals , Rats , Carotid Artery, Common , Reperfusion , Reperfusion Injury/prevention & control , Rats, Wistar , AdrenomedullinABSTRACT
The trend toward designing large hydrophobic molecules for lead optimization is often associated with poor drug-likeness and high attrition rates in drug discovery and development. Structural simplification is a powerful strategy for improving the efficiency and success rate of drug design by avoiding "molecular obesity". The structural simplification of large or complex lead compounds by truncating unnecessary groups can not only improve their synthetic accessibility but also improve their pharmacokinetic profiles, reduce side effects and so on. This review will summarize the application of structural simplification in lead optimization. Numerous case studies, particularly those involving successful examples leading to marketed drugs or drug-like candidates, will be introduced and analyzed to illustrate the design strategies and guidelines for structural simplification.
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Bronchopulmonary dysplasia (BPD) is a common respiratory disease in premature infants characterized by alveolar and pulmonary vascular dysplasia.There are currently no effective preventive and therapeutic measures.Adrenomedullin (ADM) is a vasodilator peptide produced by endothelial and smooth muscle cells of the systemic and pulmonary circulation,which promotes alveolar growth and pulmonary angiogenesis.ADM protects the respiratory system through three signaling pathways:(1) it promotes pulmonary vascular endothelial cell proliferation through extracellular regulated protein kinases (ERK);(2) attenuates lung cell apoptosis and improves cell proliferation through protein kinase B (PKB) pathway;(3) it down-regulates NADPH oxidase 4 (NOX4) expression in lung tissue to inhibit oxidative stress and improves BPD.So ADM signaling axis may be a potential therapeutic target for human infants with BPD.It provides theoretical basis for clinical prevention and treatment of BPD in premature infants.
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The Adrenomedullin peptide hormone has a potent vasodilatory activity. Nevertheless this novel peptide has exposed to be almost a ubiquitous peptide, with the many number of tissues and cell types synthesizing adrenomedullin. Adrenomedullin (ADM) and its related family peptides are calcitonin gene-related peptides (α and β-CGRPs), and intermedin/adrenomedullin-2 (IMD/ADM2) which play vital role as regulators of vascular tone and cardiovascular advances in vertebrates. Current research into their functions in reproduction has acknowledged the function of these peptides and their cognate receptors (calcitonin receptor-like receptor/receptor activity-modifying protein (CLR/RAMP) receptors) in fetal cum maternal blood circulation, feto-placental and uteroimplantation development along with that of female gamete development as well as gamete movement in the oviduct. Moreover, recent findings have enlightened the novelty, potential opportunities for the deterrence and treatment of aberrant pregnancies such as pregnancy-stimulated hypertension, preeclampsia, and IUGR. Conversely, chief efforts are still required to clarify the relationships between evident components of the CLR/RAMP signaling pathway and aberrant pregnancies before CLR/RAMP receptors can develop targets for clinical management. With this comprehension, this review summarizes current progression with specific focus on role of adrenomedullin during early implantation.
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Objective:To probe the effect of intermittent aerobic exercise on adrenomedullin secretion of kidney in hypertensive nephropathy rat (HNR),and investigate the renal mechanism for exercise in the prevention and treatment of hypertension.Method:Thirty-two HNRs were divided into two groups:intermittent exercise group (Group IAE) and control group (Group C).The IAE group performed intermittent treadmill exercise for 8 weeks and their blood pressure was measured before and after exercise by tail pressure of HNR,and serum ADM was detected by enzyme linked immunosorbent assay.Adrenomedullin (ADM) was expressed in kidney by immunohistochemistry.Quantification of specific receptor RAMP2 and CRLR in kidney ADM was detected by immunoblotting.Result:The blood pressure in the Group IAE was significantly lower than that in the Group C(P＜ 0.05).The expression of ADM of kidney and serum in the Group IAE was obviously higher than that in the Group C.The quantity of RAMP2 and CRLR in kidney was obviously increased(P ＜ 0.05).Conclusion:Intermittent aerobic exercise increases serum ADM of HNR with the up-regulated expression of ADM in kidney and their specific receptor CRLR and RAMP2.The increase of serum ADM induced by intermittent aerobic exercise may be related to the change of ADM and the regulation of its specific receptor which may be one of the mechanisms for intermittent aerobic exercise in the prevention and treatment of hypertension.
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Objective To evaluate the accuracy of serum pro-adrenomedullin (pro-ADM) concentration in predicting sepsis at different degrees of severity.Methods A total of 145 patients of both sexes,aged 18-64 yr,who were admitted to intensive care unit (ICU) of the First Hospital of Hebei Medical University from March 2015 to April 2017,with length of ICU stay＞24 h,were enrolled.The patients were divided into 3 groups according to the Sequential Organ Failure Assessment (SOFA) score within 24 h after admission to ICU:2＜SOFA score ≤ 6 mild sepsis group (n =50),6＜SOFA score ≤ 12 moderate sepsis group (n =50) and SOFA score＞ 12 severe sepsis group (n =45).Peripheral venous blood samples were collected immediately after admission to hospital for determination of serum pro-ADM concentrations by enzyme-linked immunosorbent assay.The receiver operating characteristic (ROC) curve of pro-ADM in predicting sepsis at different degrees of severity was plotted,and the area under the curve and 95％ confidence interval,cut-off value,sensitivity and specificity were calculated.Results The serum pro-ADM concentrations were significantly increased with the severity of sepsis,and the length of ICU stay was prolonged with the severity of sepsis (P＜0.05).In mild sepsis group,the area under the ROC curve was 0.770,95％ confidence interval 0.591-0.949,sensitivity 0.725,specificity 0.700 and cut-off value 6.45 nmol/L.In moderate sepsis group,the area under the ROC curve was 0.776,95％ confidence interval 0.645-0.907,sensitivity 0.813,specificity 0.760,cut-off value 8.30 nmol/L.In severe sepsis group,the area under the ROC curve was 0.83,95％ confidence interval 0.715-0.963,sensitivity 0.73,specificity 0.800,cut-off value 5.70 nmol/L.Conclusion Serum pro-ADM concentration can be used as a reliable index in predicting sepsis at different degrees of severity.
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Objective To study the correlation between intermedin 1-53 (IMD 1-53) with hypertension and formation of carotid atheromatous plaques.Methods One hundred and thirty-eight patients with 1-2 degree hypertension admitted to our hospital for carotid ultrasonography were divided into atheromatous plaques group (n =98) and atheromatous plaques-free group (n=40) with 30 subjects undergoing physical examination served as a control group in this study.Their serum IMD 1-53 level,blood pressure,and carotid IMT were measured and compared.Results The SBP [(158.57±13.55)mm Hg and (145.16±14.54)mm Hg vs (125.24±10.64)mm Hg,1 mm Hg=0.133 kPa],DBP[(95.23±5.62)mm Hg and (87.15±4.72)mm Hg vs (80.31±4.62)mm Hg] and carotid IMT[(1.26±0.38)mm and (1.05±0.28)mm vs (0.87±0.38)mm] and serum IMD 1-53 level (7.20±1.62 ng/L vs 5.05±1.85 ng/L,P＜0.01) were significantly higher in atheromatous plaques group and atheromatous plaques-free group than in control group and in atheromatous plaques group than in atheromatous plaques-free group.Conclusion IMD 1-53 is involved in the formation of carotid atheromatous plaques and can resist or delay the formation of carotid atheromatous plaques.
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Objective To preliminarily reveal the neurovascular effect of electroacupuncture and adrenomedullin (ADM) in cerebral ischemia-reperfusion injury.Methods Rat changes after middle cerebral artery ischemia and reperfusion, and the effect of electroacupuncture and ADM on them were investigated using the neurological deficit score, somatosensory evoked potentials and TTC staining technique.Results Electroacupuncture and ADM can significantly improve the neurological deficit score after cerebral ischemia and reperfusion (P0.05). The P1-N1 and N1-P2 peak values of somatosensory evoked potentials decreased significantly at 30 min after ischemia (P0.05).Conclusions Electroacupuncture can reduce neurological impairment and improve brain blood supply after cerebral ischemia and reperfusion. That is similar to the neurovascular effect of ADM.
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AIM: To explore the regulatory effect of adrenomedullin (ADM) on pulmonary oxidative stress in the rats with pulmonary hypertension induced by high blood flow.METHODS: Healthy male SD rats (n=22) were randomly divided into control group, shunt group and shunt with ADM group.Abdominal aorta and inferior vena cava shunting was produced in the rats in shunt group and shunt with ADM group.After 8 weeks, ADM (1.5 μg·kg-1·h-1) was administered into the rats in shunt with ADM group subcutaneously by mini-osmotic pump for 2 weeks.Mean pulmonary artery pressure (mPAP) was evaluated by a right cardiac catheterization procedure.The ratio of right ventricular mass to left ventricular plus interventricular septal mass [RV/(LV+SP)] and relative medial thickness (RMT) in pulmonary muscularized arteries were calculated.The content of malonaldehyde (MDA), total antioxidative capacity (T-AOC), and activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in lung tissues were detected by colorimetry.The expression of NADPH oxidase 4 (NOX4) in the lung tissue was analyzed by Western blot.RESULTS: Compared with control group, the mPAP, RV/(LV+SP) and RMT in pulmonary muscularized arteries in shunt group were all significantly increased.The content of MDA and the expression of NOX4 in the lung tissues were significantly increased.The T-AOC, and activity of SOD and GSH-Px in the lung tissues were significantly decreased.However, mPAP, RV/(LV+SP) and RMT in pulmonary muscularized arteries in shunt with ADM group were significantly decreased as compared with shunt group.Meanwhile, ADM decreased the content of MDA and the expression of NOX4 in the lung tissues, but increased the T-AOC, and activity of SOD and GSH-Px in the lung tissue of shunt rats.CONCLUSION: ADM inhibits oxidative stress response in the development of pulmonary hypertension and pulmonary vascular structural remodeling induced by high pulmonary blood flow in the rats by down-regulating the NOX4 expression and strengthening the anti-oxidation response.
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Objective To study the change of mid-regional pro-adrenomedullin(MR-proADM) level in patients with chronic heart failure and its significance.Methods Randomly selected 330 patients with chronic heart failure,including 120 cases of grade Ⅱ and Ⅲ,90 cases of grade Ⅳ,90 cases were selected.,Within 24 h after the entry of the group and 7 d after symptom relieved,serum N-terminal pro-B-type natriuretic peptide(NT-proBNP),MR-proADM,cardiac ultrasound(left ventricular diameter(LVEDD) and left ventricular function(LVEF)) were measured.Ninety physical examination in the same period as healthy control during the same period.The change of MR-proADM level in patients with chronic heart failure and its significance were analyze.Results Compared with the healthy control group,the level of MR-proADM in patients with chronic heart failure(grade Ⅱ:(641.55±106.34) pmol/l;grade Ⅲ:(684.46±101.33) pmol/l;grade Ⅳ:(737.81±479.37) pmol/l) was significantly higher than that in the control group(610.22±60.84) pmol/l),the difference was statistically significant(F=5.33,P=0.001).At the same time,the level of MR-proADM was increased with the aggravation of heart failure,after drug treatment will decline((608.09±100.81),(617.64±94.32),(642.22±163.53) pmol/L).And MR-proADM levels were significantly positively correlated with NT-proBNP levels and NYHA classification in patients with heart failure(r=0.194,P=0.003;r=0.206,P=0.000).Conclusion MR-proADM has important guiding significance in the diagnosis,classification of patients with chronic heart failure.
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Objective To explore the effect of intermedin ( IMD ) and adrenomedullin ( ADM ) on cerebral microcirculation in rats with cerebral ischemia. Methods Rat cerebral ischemia ( CI) model was established by middle cerebral artery occlusion. 40 SPF male adult Sprague-Dawley ( SD) rats were randomly divided into three groups:CI+NS ( normal saline) group, CI+ADM group and CI+IMD group, which were used to observe the changes of brain surface microcirculatory perfusion with a laser Doppler flowmeter. Results The differences of brain surface microcirculatory perfusion were statistically significant among the CI+NS group, CI+ADM group and CI+IMD group ( F=53. 426, P<0. 05 ) . Multiple comparison showed that the brain surface microcirculatory perfusion in the CI+IMD group was higher than that of the CI+NS group and CI+ADM group. Conclusions Intermedin can improve the cerebral microcirculation in rats with cerebral ischemia, and its therapeutic effect is better than adrenomedullin.
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La adrenomedulina (AM) es un péptido involucrado en la regulación cardiovascular. En el cerebelo, la densidad de los receptores de la AM se encuentra alterada durante la hipertensión, sugiriendo un posible papel del sistema adrenomedulinérgico cerebelar en la regulación de la presión arterial (PA). El objetivo del presente estudio fue evaluar el efecto funcional in vivo de la AM durante el estrés agudo, mediante la administración in situ de AM en el vermis cerebelar de la rata. Se emplearon ratas adultas normotensas Wistar Kyoto (WKY) y Sprague Dawley (SD) y ratas espontáneamente hipertensas (SHR) las cuales fueron anestesiadas y posteriormente canuladas en el vermis cerebelar. El estrés se indujo mediante el uso del estímulo eléctrico plantar (EEP). Los animales fueron divididos en grupos que recibieron AM (0,2 o 200 pmol/5μL) o vehículo (solución fisiológica, 5μL). La PA se determinó antes del experimento y después de la administración del tratamiento respectivo, seguida de la aplicación del EEP (100 V, 5 Hz, 10 mseg, durante 4 minutos). La PA se determinó mediante pletismografía digital no invasiva. Los resultados demuestran que la microinyección de AM (0,2 y 200 pmol) in situ en el vermis cerebeloso en ratas SD, WKY y SHR disminuye significativamente la respuesta presora frente al estrés inducido por el EEP, lo que sugiere que la acción hipotensora está mediada a través de la regulación del eflujo simpático. Estos hallazgos demuestran la participación de la AM cerebelosa en la regulación de la respuesta cardiovascu lar frente al estrés.
Adrenomedullin (AM) is a peptide involved in cardiovascular regulation. In the cerebellum, the density of AM receptors is altered during hypertension, suggesting a pos sible role of cerebellar adrenomedulinergic system in the regulation of blood pressure (BP). The aim of this study was to evaluate the functional role of AM during acute stress, by in situ administration of AM into the cerebellar vermis in rats. Adult normotensive Wistar Kyoto (WKY) and Sprague Dawley (SD) rats and spontaneously hypertensive rats (SHR), were anes thetized and their cerebellar vermis cannulated. Footshock was used as stressor. Animals were divided into groups that received either AM (0.2 and 200 pmol/5μL) or vehicle (physiological saline, 5μL). The BP was determined, using noninvasive digital plethysmography, before and after treatment, followed by footshock (100V, 5 Hz, 10 msec, for 4 minutes). The results show that microinjection of AM (0.2 and 200 pmol) in situ into the cerebellar vermis in SD, WKY and SHR rats, significantly decreased the pressor response induced by footshock stress, sugges ting that the hypotensive action is mediated through regulation of sympathetic outflow. Taken together, our results demonstrate a role of cerebellar AM in the regulation of cardiovascular response to stress.
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Animals , Male , Stress, Physiological/physiology , Cerebellum/physiology , Adrenomedullin/physiology , Rats, Wistar , Rats, Sprague-DawleyABSTRACT
La adrenomedulina (ADM), péptido vasodilatador de 52 aminoácidos, ubícuo, posee múltiples efectos fisiológicos que contribuyen a respuestas homeostáticas. Se encuentra en concentraciones importantes en la glándula suprarrenal, el pulmón, el sistema cardiovascular y el renal. Ejerce acciones biológicas a través de sus receptores AM1 y AM2, heterodímeros constituidos por el receptor semejante al receptor de calcitonina (CLR) y proteínas modificadoras de la actividad del receptor (RAMPs), CLR/RAMP2 y CLR/RAMP3, respectivamente. La principal vía de señalización es la adenilato ciclasa/AMPc en patologías cardiovasculares y renales, así como en la sepsis. Los niveles plasmáticos de ADM se elevan proporcionalmente con el incremento de la presión sanguínea y con el grado de daño renal, en pacientes con hipertensión arterial; así mismo, dichos niveles se correlacionan con el grado de hipertrofia cardíaca y arterial. La ADM tiene efectos renales, tubulares, ya que tiene acción diurética y natriurética; posee efectos vasodilatadores que producen aumento de la tasa de filtración glomerular y del flujo sanguíneo renal. Este péptido inhibe la proliferación y la generación de radicales libres en células mesangiales; actúa inhibiendo la secreción de aldosterona en la zona glomerulosa y de endotelina-1 en células musculares lisas vasculares. Se ha propuesto como marcador de riesgo en diversas patologías, especialmente en la insuficiencia renal crónica; en esta patología, que cursa con hipertrofia compensadora de los glomérulos y proliferación del mesangio, la administración de ADM disminuye los niveles de proteinuria, lo que sugiere que además de un importante rol modulador en la presión arterial, pudiera ser una opción terapéutica para la insuficiencia renal crónica.
Adrenomedullin (AM) is a potent vasodilatory 52-aminoacid peptide hormone, ubiquitous with multiple physiological effects which contribute to homeostatic responses. Significantly, it is distributed in the adrenal gland, lung, cardiovascular and renal system. The biological effects of AM are directly mediated by specific receptors as heterodimers composed of the calcitonin-receptor-like receptor (CLR) and one of two receptor activity modifying proteins (RAMP2 or RAMP3). The CLR/RAMP2 (AM1 receptor) is more highly AM-specific than The CLR/RAMP3 (AM2 receptor). Plasma levels of AM are elevated proportionately to the increase in blood pressure and degree of renal damage in patients with hypertension; likewise, these levels are correlated with the degree of heart and arterial hypertrophy. AM has renal vasodilatory, natriuretic and diuretic actions; increased glomerular filtration rate and renal blood flow. AM inhibits proliferation and reactive oxygen species generation in mesangial cells; also inhibits aldosterone secretion in the zona glomerulosa and endothelin-1 in vascular smooth muscle cells. Therefore, it is proposed as a new marker in various diseases, especially chronic renal failure. This disease presents compensatory hypertrophy of the glomeruli and mesangial proliferation, administration of AM reduces the levels of proteinuria, suggesting that AM has an important modulator role in blood pressure and could be a therapeutic option for chronic renal failure.
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Humans , Adrenomedullin/physiology , Kidney/physiology , Adrenomedullin/therapeutic use , Kidney/physiopathology , Kidney Failure, Chronic/drug therapyABSTRACT
Objective To investigate the regulatory effect of adrenomedullin (AM) on the cell-cell contact formation of podocytes and the possible mechanism.Methods Podocytes were treated with AM (10-7 mol/L),AM combined with a PKA inhibitor H89 (10-4 mol/L),and forskolin (10-5 mol/L) as positive control respectively for 12 hours.Immunofluorescent staining was applied to observe the distribution of cell adhesion molecules and actin-associated proteins.Western blotting assay was used to assess their protein levels.Rho GTPases activity was analyzed by GST-pull down assay and their protein levels were tested by Western blotting.Results AM induced the redistribution of adhesion molecules,actin-associated proteins as well as the F-actin at cell-cell contacts between podocytes.This effect was similar to that of forskolin and could be blocked by H89.The levels of those proteins did not change significantly (P ＞ 0.05).AM up-regulated the activities of RhoA,Rac1 and Cdc42 (P ＜ 0.05),which were partially blocked by H89.The protein levels of Rho GTPases showed no difference compared with the control (P ＞ 0.05).Conclusions AM may promote cell-cell contact formation of podocytes,probably through enhancing the activity of Rho GTPases and then resulting in the redistribution of adhesion molecules,actin-associated proteins and F-actin,which is partially mediated through cAMP-PKA signaling pathway.
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ABSTRACT:Objective To study the expressions of adrenomedullin (ADM)andβ-catenin in human osteosarcoma of different differentiation grading and their clinical significance.Methods We chose 33 cases of human osteosarcoma with different pathological grading (14 cases of high differentiation grading and 1 9 cases of low grading)and 10 cases of human osteochondroma to detect the expressions of ADM protein,total (T)β-catenin and phosphorylated (P)β-catenin using immunohistochemical method. Results The expression of ADM was detected in the cytoplasm.In osteosarcomas of higher and lower differentiation grading,the positive rate of ADM was 57.1% (8/14)and 100% (1 9/1 9),respectively,with significant differences (P =0.0030.05).The expression of P-β-catenin was detected in the cytoplasm.In osteosarcomas of higher and lower differentiation grading,the positive rate of P-β-catenin was 35.7% (5/14)and 78.9% (1 5/1 9),with a significant difference (P =0.029 0.05 )or between T-β-catenin and P-β-catenin (P = 0.052 > 0.05 ).Conclusion The expressions of ADM and P-β-catenin proteins and the different distributions of expression of T-β-catenin protein in cytoplasm and/or nucleus are important indicators for judging the pathological grading of osteo-sarcoma.
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Objective To explore the relationship between preoperative serum pro-adrenomedullin (pro-ADM) in the patients with closed rupture of small intestine,and postoperative secondary abdominal infection,and to study the value of postoperative serum pro-ADM in assessment of severity of postoperative secondary abdominal infection of this patients.Methods Eighty-five patients with closed rupture of small intestine treated in the emergency surgery from June 2014 to May 2015 were selected.According to the presence of postoperative abdominal infection or not,these patients were divided into infection group and non-infection group.The infection group was further divided into SIRS,sepsis,severe sepsis,septic shock subgroups as per the severity of infection.The levels of serum pro-ADM,interleukin-6 (IL-6),C-reactive protein (CRP) were determined and the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score was calculated at admission and the first day、the second day and the third day after operation.Comparisons of these biomarkers were carried out to find out the factors associated with postoperative abdominal infection.In addition,ROC curve was used to verify the factors for predicting the abdominal infection of these patients after operation.The relationship between serum levels of pro-ADM and APACHE Ⅱ score after operation was analyzed.The severity of abdominal infection after operation was assessed with laboratory findings.Results Compared with non-infection group,preoperative and postoperative serum pro-ADM (P =0.03,P ＜ 0.01),IL-6 (P =0.02,P ＜0.01) levels and APACHE Ⅱ scores (P ＜ 0.01,P ＜ 0.01) were significantly higher in infection group (P ＜ 0.05).In the infection groups,the postoperative levels of serum pro-ADM and APACHE Ⅱ scores were increased with the severity of infection increased (r =0.924),and the difference between the groups was statistically significant (P ＜ 0.05),but there were no significant differences in IL-6 and CRP levels between two groups.Conclusion (1) The serum levels of pro-ADM before operation has value in predicting the genesis of abdominal infection in these patients after operation.(2) The serum levels of pro-ADM after operation has value in severity assessment of abdominal infection in these patients after operation.
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Objective To investigate the effect of adrenomedullin on epithelial barrier function of Caco-2 cells under hypoxia/reoxygen-ation injury and the molecular mechanism.Methods Build hypoxia/reoxygenation injury model,Caco-2 cells were randomly divided into three groups:normal control group( N) ,hypoxia/reoxygenation group( H/R) and hypoxia/reoxygenation plus 1 μmol/L adrenomedullin group(H/R+AM).Transepithelial electrical resistance(TEER) was determined,and the laser confocal microscope was used to detect the construction of tight junction.The protein expressions of tight junction proteins (Occludin and ZO-1) and nuclear factor kappa B(NF-κBp65) were examined by using western blotting.Results Pretreatment of AM significantly attenuated the disruption of morphological structure of tight junction caused by H/R.The TEER of N group,H/R group and H/R+AM group were(157.68 ±5.54),(96.06 ±3.42),(134.56 ± 4.72) Ohm/cm2 respectively.After AM pretreatment, the TEER were significantly raised compared with that in H/R group by 40.00%(P<0.05).The relative protein expressions of Occludin and ZO-1 in N group,H/R group and H/R+AM group were [(0.43 ±0.03), (0.27 ±0.04)],[(0.20 ±0.03),(0.15 ±0.07)],[(0.32 ±0.15),(0.21 ±0.03)]respectively.After AM pretreatment,the protein ex-pression of Occludin and ZO-1 were significantly raised compared with that in H/R group(by 60.00% and 40.00%).Simultaneously,the relative protein expressions of NF-κBp65 in N group,H/R group and H/R+AM group were (0.53 ±0.30),(2.89 ±0.16),(1.75 ±0.25) respectively.And H/R induced NF-κBp65 activation was significantly inhibited by AM treatment (decreased as compared with H/R group by 40.00%).Conclusion AM can attenuate intestinal epithelial barrier dysfunction by regulating the expression of intestinal tight junction pro-teins.The inhibition of NF-κB activation may be involved in this mechanism.
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Objective To observe the effect of silencing the expression of ADM using RNA interfering technique on the expression ofβ-catenin and terminal differentiation of osteosarcoma cells.Methods After the intervention of 0 h,48 h and 72 h by ADM siRNA,we observed the change of distribution ofβ-catenin in F5M2 using immunocytochemistry staining.The expression levels of total and phosphorylatedβ-catenin and GSK3βwere detected by Western blot after the intervention by ADM siRNA.The F5 M2 cells treated with ADM siRNA were subjected to HE staining,alkaline phosphatase (ALP)assay and immunocytochemistry staining to investigate the biological effects of ADM siRNA on the morphology and terminal differentiation of F5M2 cells.Results After the intervention of 0 h,48 h and 72 h by ADM siRNA,the distribution ofβ-catenin was transferred from the cytoplasm to the nucleus.ADM siRNA downregulated the expression level of P-β-catenin and upregulated P-GSK3βdetected by Western blot.HE staining revealed that the configuration of F5M2 had undergone restorational changes similar to those of normal cells after ADM siRNA treatment.ALP assay and immunocytochemistry staining showed that the expression of the earlier and later molecular biomarkers of terminal differentiation,including ALP and osteocalcin were strongly positive.Conclusion Silencing the expression of ADM can activateβ-catenin to transfer to the nucleus from the cytoplasm,and induce osteosarcoma cells to make terminal differentiation through activatingβ-catenin signaling pathway.