Role of the NF-kB/parkin/vegfr-1 pathway associated with hypoxic-ischemic insult in germinal matrix samples of newborn infants / O papel da via NF-kB/parkin/vegfr-1 associada à agressão hipóxico-isquêmica em amostras de matriz germinativa de pacientes recém-nascidos

Abstract Objective: Given the high proliferative activity of germinal matrix and its direct correlation with hypoxemia, it is necessary to investigate the possible molecular regulation pathways, to understand the existing clinical relationship between the hypoxic-ischemic insult and the biomarkers NF-kB, AKT-3, Parkin, TRK-C and VEGFR-1. Methods: A hundred and eighteen germinal matrix samples of the central nervous system of patients who died in the first 28 days of life were submitted to histological and immunohistochemistry analysis to identify the tissue immunoexpression of those biomarkers related to asphyxia, prematurity, and death events within 24h. Results: A significantly increased tissue immunoexpression of NF-kB, AKT-3 and Parkin was observed in the germinal matrix of preterm infants. In addition, significantly decreased tissue immunoexpression of VEGFR-1 and NF-kB was observed in patients who experienced asphyxia followed by death within 24 hours. Conclusions: The results suggest a direct involvement between the hypoxic-ischemic insult and NF-kB and VEGFR-1 markers since a decreased immunoexpression of these biomarkers was observed in asphyxiated patients. Furthermore, it is suggested that there was not enough time for VEGFR-1 to be transcribed, translated and expressed on the surface of the plasma membrane. This temporality can be observed in the relationship between NF-kB expression and the survival time of individuals who died within 24 hours, suggesting that this factor is essential for the production of VEGFR-1 and, therefore, to carry out the necessary remodeling effect to neovascularize the affected region.

RESUMO Objetivo: Dada a alta atividade proliferativa da matriz germinativa e sua correlação direta com a hipoxemia, é necessário investigar as possíveis vias de regulação molecular para entender a relação clínica existente entre o insulto hipóxico-isquêmico e os biomarcadores NF-kB, AKT -3, Parkina, TRK-C e VEGFR-1. Métodos: Cento e dezoito amostras de matriz germinativa do sistema nervoso central de pacientes que faleceram nos primeiros 28 dias de vida foram submetidas a análise histológica e imuno-histoquímica para identificar a imunoexpressão tecidual desses biomarcadores relacionados a eventos de asfixia, prematuridade e óbito em 24 horas. Resultados: Observou-se uma imunoexpressão tecidual significativamente aumentada de NF-kB, AKT-3 e Parkin na matriz germinativa de prematuros. Além disso, constatou-se uma imunoexpressão tecidual significativamente diminuída de VEGFR-1 e de NF-kB em pacientes que apresentaram asfixia seguida de morte em 24 horas. Conclusões: Os resultados sugerem o envolvimento direto entre o insulto hipóxico-isquêmico e os marcadores NF-kB e VEGFR-1, visto que se observou uma imunoexpressão diminuída destes biomarcadores nos pacientes asfixiados. Além disso, sugere-se que não houve tempo suficiente para que o VEGFR-1 fosse transcrito, traduzido e expresso na superfície da membrana plasmática. Essa temporalidade pode ser observada na relação entre a expressão de NF-kB e o tempo de vida dos indivíduos que morreram em 24 horas, o que sugere que esse fator é essencial para a produção do VEGFR-1 e, portanto, para realizar o efeito remodelador necessário para neovascularizar a região afetada.

Reducing neuroinflammation and improving neurobehavioral functions: the protective roles of resveratrol for germinal matrix hemorrhage in neonatal rat model / 中国新生儿科杂志

Objective:To study the roles of resveratrol in reducing neuroinflammation and improving neurobehavioral functions after germinal matrix hemorrhage (GMH) in neonatal rat model.Methods:GMH model was established intraparenchymally injecting bacterial collagenase in 7-day-old SD rats. 108 rats were randomly assigned into 18 groups (6 in each group), including 4 sham groups, GMH (12 h, 24 h, 72 h, 7 d) groups, 3 GMH+vehicle (dimethylsulfoxide, DMSO) groups, 5 GMH+resveratrol (10 mg/kg, 100 mg/kg, 1 000 mg/kg) groups and 2 GMH+resveratrol+EX527 (SIRT1 inhibitor) groups. Negative geotaxis and righting reflex tests were used to evaluate the short-term neurobehavior. Water maze, foot fault and Rotor-Rod tests were used to assess the long-term neurobehavior. Immunofluorescence was used to quantify the IL-1β and MPO positive cells (inflammatory markers) in peri-hematoma area. Western blot was used to evaluate the expression of relevant proteins in the brain.Results:Endogenous sirtuin-1(SIRT1) decreased to the lowest level at 24 h and then increased gradually. Phosphorylated NF-κB increased at 12 h, peaked at 72 h and returned to normal level at 7 d after GMH. Compared with the control group and other doses groups, GMH treated with resveratrol (100 mg/kg) had higher short-term behavioral scores at 48 h and 72 h. Compared with the control group, the resveratrol (100 mg/kg) group also had higher scores in water maze, foot fault and Rotor-Rod tests 22 days later. Immunofluorescence showed less positive IL-1β and MPO cells around hematoma in GMH+resveratrol group than both GMH+vehicle group and GMH+resveratrol+EX527 group. Western blot indicated that IL-1, TNF-α and IL-6 expressions were decreased in GMH+resveratrol group and Ex527 could offset the effects of resveratrol.Conclusions:Resveratrol (optimal dose: 100 mg/kg) can improve the short-term and long-term neurobehavioral functions of neonatal GMH rats. It can reduce GMH cells with positive inflammatory markers around the hematoma, possibly via inhibition of the SIRT1/NF-κB pathway. Resveratrol may be promising for the treatment of GMH patients.

Stimulation of mGluR5 by VU0360172 protected against germinal matrix hemorrhage in neonatal rats / 中国药理学通报

Aim To investigate the protective effect of mGluR5 activated by VU0360172 on germinal matrix hemorrhage in neonatal rats.Methods Seven day- old SD rats were randomly divided into Sham, GMH, and low-, medium-, and high-dose groups.The model was established by intracerebral injection of collagenase W-S.Then three doses of VU0360172 were injected intraperitoneal

Mecanismos de lesión cerebral en niños prematuros / Mechanisms of brain injury of the premature baby

Los nacimientos prematuros son uno de los principales indicadores de salud de un país. Están asociados a una alta mortalidad e importante morbilidad en niños con parálisis cerebral y otros trastornos del neurodesarrollo, incluyendo problemas cognitivos y del aprendizaje. Los principales tipos de lesión encefálica en los recién nacidos prematuros son: a) las lesiones de la sustancia blanca, generalmente asociadas a alteraciones neuronales y axonales en la corteza cerebral y otras zonas de sustancia gris; b) hemorragias intracraneanas que incluyen las de la matriz germinal, intraventriculares e intraparenquimatosas y c) del cerebelo. Las lesiones de sustancia blanca incluyen la leucomalacia periventricular quística, no quística (con focos de necrosis microscópicos) y lesiones difusas de sustancia blanca, no necróticas. Estas lesiones tienen múltiples factores etiológicos. Las características anatómicas y fisiológicas de las estructuras vasculares periventriculares predisponen a la sustancia blanca a ser muy vulnerable a las situaciones de isquemia cerebral y, en interacción con factores infecciosos/inflamatorios, activan a las microglías generando estrés oxidativo (por liberación de radicales libres del oxígeno y del nitrógeno), liberación de citoquinas proinflamatorias, liberación de glutamato, fallo energético y alteración de la integridad vascular. Todo lo anteriormente mencionado genera una particular vulnerabilidad de los pre-oligodendrocitos que termina alterando la mielinización. La hipoxia-isquemia también puede producir necrosis neuronal selectiva en diferentes regiones encefálicas. La matriz germinal es un área altamente vascularizada en la región subependimaria periventricular con una estructura capilar muy frágil que la predispone a las hemorragias.

Preterm birth is one of the main country health indicators. It is associated with high mortality and significant morbidity in preterm newborns with cerebral palsy and potential long-term neurodevelopmental disabilities like cognitive and learning problems. The main lesions could be: a) white matter injuries, generally associated with cortical and other regions of grey matter neuronal-axonal disturbances; b) intracranial hemorrhage that includes germinal matrix, intraventricular and parenchymal, c) cerebellum injuries. The white matter lesions include cystic and non-cystic (with microscopic focal necrosis) periventricular leukomalacia and non-necrotic diffuse white matter injury. Multiple etiologic factors are associated with these injuries. Anatomical and physiological characteristics of periventricular vascular structures predispose white matter to cerebral ischemia and, interacting with infection/inflammation factors, activate microglia, generating oxidative stress (mediated by free oxygen and nitrogen radicals), pro-inflammatory cytokine and glutamate toxicity, energetic failure and vascular integrity disturbances. All these factors lead to a particular vulnerability of pre-oligodendrocytes that will affect myelination. Hypoxia-ischemia also may produce selective neuronal necrosis in different cerebral regions. Germinal matrix is a highly vascularized zone beneath ependymal or periventricular region that constitutes a capillary bed with a particular structural fragility that predispose it to hemorrhage.

Trans-fontanellar ultrasound evaluation of neonates and infants in a tertiary care hospital in Garhwal referred for sonography

Ultrasound is widely used to diagnose various conditions in neonates and infants like intracranial hemorrhage, hypoxic-ischemic insult, changes of birth asphyxia and congenital disorders of the ain like hydrocephalus, etc as it is easily available, simple and noninvasive and more important is that it lacks ionizing radiation. The present hospital-based study of cranial sonography (Trans-fontanellar ultrasound) done in the Department of Radiodiagnosis, HNB Base government teaching hospital situated in Srinagar Garhwal, was done with the aim of assessing the burden of intracranial disorders in neonates and infants, especially the premature newborns who get admitted in neonatal intensive care units. A retrospective descriptive analysis was done on the data of 196 CUSG done.70.4 % CUSG were performed on neonates, 24.5% were performed on age group 29 days to 6 month age group and rest on age more than 6 months. The male-female gender ratio was 1: 0.96. The most common neonatal pathological conditions were changes of Hypoxic- ischemic encephalopathy (HIE) (28.4 %). Cranial sonography is a useful diagnostic tool in the evaluation of neonatal and infant ain and is a well-established mode of making primary diagnosis.It is preferred because it can be rapidly performed and is a portable, cost-effective test.

Effect of hyaluronidase on expression of neuron-glial antigen 2 and myelin basic protein after germinal matrix-intraventricular hemorrhage in premature rabbits / 中华实用儿科临床杂志

Objective@#To investigate the neuroprotective effect of hyaluronidase(HAase) on rabbit brain tissue in germinal matrix-intraventricular hemorrhage(GM-IVH) premature rabbits.@*Methods@#Eighty premature rabbits of gestation age 29 days were randomly divided into normal group, GM-IVH control group and HAase treatment group.The rabbits in GM-IVH control group and HAase treatment group were given intraperitoneal injection of 50 g/L glycerol to establish GM-IVH animal model, while the premature rabbits in normal group were given the same dose of 9 g/L saline.The GM-IVH was screened by using cranial ultrasound.The premature rabbits in HAase treatment group were given HAase into the lateral forebrain while the rabbits in the GM-IVH control group were given the same dose of 9 g/L saline.In 3 days, 7 days and 14 days after birth, the premature rabbits were killed and brain tissue were separated.The expression of neuron-glial antigen 2(NG2) were detected by adopting immunohistochemical method.The expression of myelin basic protein(MBP) were detected by Western blot method.@*Results@#The expression of NG2 in 3 groups of premature rabbits decreased gradually with the increase of age[the values of NG2 in the normal group were(62.65±33.58)×104, (15.61±4.22)×104, and (13.54±4.51)×104 on the 3rd, 7th, and 14th day; those in the GM-IVH control group were (54.58±25.48)×104, (48.91±22.49)×104, (7.18±2.28)×104 on the 3rd, 7th, and 14th day; those in the HAase treatment group were (148.13±27.30)×104, (88.38±14.55)×104, (77.98±18.96)×104 on the 3rd, 7th, and 14th day]. At the same time, the expression of NG2 in the HAase treatment group was higher than that of the GM-IVH control group, and the differences were statistically significant among the 3 groups at any time(all P<0.05). The expression of MBP protein in the 3 groups increased with the increase of day age[MBP protein in the normal group were (0.30±0.22)×103, (1.91±0.43)×103, and (5.67±2.14) ×103 on the 3rd, 7th, and 14th day; those in the GM-IVH control group were (0.87±0.12)×103, (1.15±0.22)×103 and (2.54±0.69) ×103 on the 3rd, 7th, and 14th day; those in the HAase treatment group were (0.91±1.01)×103, (2.25±0.66)×103, (3.40±1.10)×103 on the 3rd, 7th, and 14th day]. The difference was statistically significant between the normal group and the HAase treatment group(P<0.05).@*Conclusions@#After HAase treatment, the expression of NG2 and MBP can be improved to a certain extent, promoting the development of oligodendrocytes and regeneration of myelin sheath.

Effect of hyaluronidase on expression of neuron＿glial antigen 2 and myelin basic protein after germinal ma-trix＿intraventricular hemorrhage in premature rabbits / 中华实用儿科临床杂志

Objective To investigate the neuroprotective effect of hyaluronidase( HAase)on rabbit brain tissue in germinal matrix-intraventricular hemorrhage( GM -IVH)premature rabbits. Methods Eighty premature rabbits of gestation age 29 days were randomly divided into normal group,GM-IVH control group and HAase treatment group. The rabbits in GM -IVH control group and HAase treatment group were given intraperitoneal injection of 50 g/L glycerol to establish GM-IVH animal model,while the premature rabbits in normal group were given the same dose of 9 g/L saline. The GM-IVH was screened by using cranial ultrasound. The premature rabbits in HAase treat-ment group were given HAase into the lateral forebrain while the rabbits in the GM-IVH control group were given the same dose of 9 g/L saline. In 3 days,7 days and 14 days after birth,the premature rabbits were killed and brain tissue were separated. The expression of neuron - glial antigen 2( NG2 )were detected by adopting immunohistochemical method. The expression of myelin basic protei(n MBP)were detected by Western blot method. Results The expression of NG2 in 3 groups of premature rabbits decreased gradually with the increase of ag[e the values of NG2 in the normal group were(62. 65 ± 33. 58)×104(,15. 61 ± 4. 22)×104 ,and(13. 54 ± 4. 51)×104 on the 3rd ,7th ,and 14th day;those in the GM-IVH control group were(54. 58 ± 25. 48)×104 ,(48. 91 ± 22. 49)×104 ,(7. 18 ± 2. 28)×104 on the 3rd ,7th , and 14th day;those in the HAase treatment group were(148. 13 ± 27. 30)×104 ,(88. 38 ± 14. 55)×104 ,(77. 98 ± 18. 96)×104 on the 3rd,7th,and 14th day]. At the same time,the expression of NG2 in the HAase treatment group was higher than that of the GM-IVH control group,and the differences were statistically significant among the 3 groups at any time(all P<0. 05). The expression of MBP protein in the 3 groups increased with the increase of day age[MBP protein in the normal group were(0. 30 ± 0. 22)×103 ,(1. 91 ± 0. 43)×103 ,and(5. 67 ± 2. 14)×103 on the 3rd , 7th,and 14th day;those in the GM-IVH control group were(0. 87 ± 0. 12)×103,(1. 15 ± 0. 22)×103 and(2. 54 ± 0. 69)×103 on the 3rd,7th,and 14th day;those in the HAase treatment group were(0. 91 ± 1. 01)×103 ,(2. 25 ± 0. 66)×103 ,(3. 40 ± 1. 10)×103 on the 3rd,7th ,and 14th day]. The difference was statistically significant between the normal group and the HAase treatment grou(p P<0. 05). Conclusions After HAase treatment,the expression of NG2 and MBP can be improved to a certain extent,promoting the development of oligodendrocytes and regeneration of myelin sheath.

Effect of thyroid hormone on the expression of deiodinase and myelin basic protein in brain tissue of preterm rabbit with germinal matrix-intraventricular hemorrhage / 临床儿科杂志

ObjectivesTo investigate the effect of thyroid hormone on the expression of deiodinase and myelin basic protein in brain tissue of neonatal rats with germinal matrix-intraventricular hemorrhage (GM-IVH), and its nerve protective effect.Methods Sixty premature rabbits of gestational age 29 days were randomly divided into control, model and intervention groups. The prema-ture rabbits in model and intervention groups were given intraperitoneal injection of 50% glycerol to establish GM-IVH animal model and the premature rabbits in control group were given the same dose of saline. The GM-IVH was screened by cranial ultrasound. The premature rabbits with GM-IVH in intervention group were given intraperitoneal injection of triiodothyronine and the other two groups were given the same dose of saline. At 3 days, 7 days and 14 days after birth, the premature rabbits were killed and the brain tissue were quickly separated. The expression of type 2 iodothyronine deiodinase (D2) and type 3 iodothyronine deiodinase (D3) of periventricular zone tissue were detected by western blot method. The expression of myelin basic protein (MBP) were detected by immunohistochem-ical method.ResultsThe expression of D2 in brain tissue increased gradually in three groups. At the same time point, the expression of D2 was the lowest in the model group, the highest in the control group, and the middle in the intervention group. The differences of D2 among three groups were statistically signiifcant (allP<0.001). The expression of D3 increased gradually in the control and model groups, while it was the highest in the intervention group at day 7. At the same time point, the expression of D3 in the control group was the lowest, it was higher in intervention group than that in the model group at 3 days, then it was higher in the model group than that in the intervention group. The differences of D3 among three groups were statistically signiifcant (allP<0.001). The expression of MBP increased gradually in the control and intervention groups, and it decreased gradually in the model group. At the same time point, the MBP was the lowest in the model group, it was the highest in the control group, and it was the middle in the intervention group. The differences of MBP among three groups were statistically signiifcant (P<0.001).ConclusionsThe expressions of D2 and MBP in the brain tissues of premature rabbit with the GM-IVH increased while the expression of D3 decreased in some extent after thyroxine intervention.

Risk Factors Associated with Germinal Matrix-Intraventricular Hemorrhage in Preterm Neonates

OBJECTIVE: The purpose of this study is to identify the risk factors associated with the development of germinal matrix-intraventricular hemorrhage (GM-IVH) and the relationship of the severity of disease and prematurity. METHODS: A total of 168 premature neonates whose birth weight < or =1500 g or gestational age < or =34 weeks were examined by cranial ultrasound (CUS) for detection of GM-IVH among the babies admitted between January 2011 and December 2012 in our medical center neonatal intensive care unit. The babies were divided into two groups : GM-IVH and non-IVH. Clinical presentations, precipitating factors of the patients and maternal factors were analyzed. RESULTS: In univariate analysis, gestational age, birth weight, delivery method, presence of premature rupture of membrane (PROM) and level of sodium and glucose were statistically meaningful factors (p<0.05). But only two factors, gestational age and presence of patent ductus arteriosus (PDA) were statistically meaningful in multivariate logistic regression (p<0.05). Delivery method [normal vaginal delivery (NVD) to Caeserean section] was borderline significant (p<0.10). CONCLUSION: Presence of PDA and gestational age were the important risk factors associated with development of GM-IVH.

Progress of germinal matrix intraventricular hemorrhage in the premature infant / 国际儿科学杂志

Of all types of intracranial hemorrhage (ICH) in the neonates, germinal matrix intraventricular hemorrhage(GM-IVH) in the premature infant is the most common type, mainly attributed to the existence of immature germinal matrix, lt's usually lack of visible and specific symptoms and signs in the nervous system, so its early and final diagnosis depends on the imaging examine. GM-IVH can cause periventricular hemorrhagic infarction, post-hemorrhagic hydrocephalus, periventricular leukomalacia and the associated cerebellar hemorrhagic injury, which are critical determinants of neonatal morbidity, mortality, and neurodevelopmental outcome in the survivors. The overall aim of this article is to review the current knowledge of the cause,mechanisms, imaging diagnosis, complication, management and outcome of GM-IVH in the preterm infant.

Clinical Features of Congenital Cytomegalovirus Infection in Neonates / 대한주산의학회잡지

OBJECTIVE: Cytomegalovirus(CMV) is the most common cause of congenital infection with diverse clinical features according to the age of hosts. This study was done to evaluate clinical manifestations and complications in congenital CMV infection. METHODS: Between October 2000 and September 2001, 10 cases of congenital CMV infection were diagnosed by urine culture within 2 weeks after birth. Demographic data, clinical and laboratory findings were collected from maternal and newborn hospital records. RESULTS: There were 7 males and 3 females. Average gestational age was 38+0 weeks(34+2-41+1), average birth weight was 2462+/-598.2g(1670-3360) and 5 cases were small for gestational age. Average head circumference was 31.9+/-1.8cm(29-34.3). There were 4 cases of microcephaly, 6 cases of petechiae, 3 cases of leukocytopenia, 6 cases of thrombocytopenia. There was no case showing hepatomegaly, hepatic dysfunction, or pathologic jaundice. 4 cases showed physiologic jaundice. IgM level was elevated in 5 out of 9 cases. Polymerase chain reaction(PCR) was positive in 7 cases. All cases showed germinal matrix hemorrhage and septated cysts on cranial ultrasonography. 1 case showed cerebral calcification. 1 case showed bilateral sensorineural hearing loss on brainstem auditory evoked potentials(BAEP). There was no case showing chorioretinitis on fundoscopy. CONCLUSION: There were diverse clinical manifestations in CMV infection in neonates. All cases showed germinal matrix hemorrhage and septated cysts. When we find these findings, we should consider the possibility of CMV infection. One patient developed bilateral sensorineural hearing loss, so its early detection and follow-up might be necessary.

Estudio de factores de riesgo para la hemorragia de la matriz del prematuro / Risk factors for germinal matrix hemorrage in preterm infants

Con el propósito de facilitar el diagnóstico clínico de la hemorragia de la matriz germinal del prematuro, se diseñó un estudio de casos (n: 56) y controles (n: 66) para determinar los factores de riesgo de la madre y del niño; también se buscó asociación entre dichos factores y la gravedad del evento. No se halló relación entre los factores de riesgo maternos y la presencia o gravedad de la hemorragia. En cuanto a los niños, la edad gestacional menor de 31 semanas, el uso de ventilación mecánica y la persistencia del ductus arterioso mostraron una relación estadística con el evento (p < 0.05), mientras que la sumatoria de tres o más riesgos la tuvo con la gravedad del mismo (p < 0.05). Se concluye que los mencionados factores de riesgo pueden ser indicativos del desarrollo de la hemorragia de la matriz germinal en el prematuro

To facilitate clinical diagnosis of germinal matrix hemorrhage In preterm infants, a study of cases {n: 56) and controls {n: 66) was carried out. Association was investigated between hemorrhage and maternal and neonatal risk factors; also included was the correlation between such factors and seriousness of the hemorrhagic episode. No correlation was found between maternal risk factors and hemorrhage or its seriousness. Concerning neonatal risks, gestational age under 31 weeks, the use of mechanical ventilation and persistence of ductus arteriosus, independently, showed statistical correlation with the hemorrhage {p < 0.05), whereas the simultaneous presence of three or more risks correlated with intensity of hemorrhage {p < 0.05). We conclude that the abovementioned neonatal risk factors can be suggestive of the development of germinal matrix hemorrhage in preterm Infants