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The impact of NgR specific siRNA on neurite outgrowth in neonatal rats after hypoxic-ischemic brain damage

Gangming XIAO ; Xiaoqing QIN ; Xiaoguang ZHOU

Chinese Journal of Neonatology.2015;30(3):224-228. doi:10.3969/j.issn.1673-6710.2015.03.020

Objective To study the effect of chemically synthesized Nogo-66 receptor ( NgR ) specific small interfering RNA ( siRNA) on nerve regeneration and function of newborn rats after hypoxic-ischemic brain damage ( HIBD) . Methods A total of 50 HIBD newborn rats were set up. They were randomly assigned OR allocated into siNgR group ( n=20 ) , normal saline ( NS ) group ( n=20 ) and HIBD group ( n=10 ) . The rats of siNgR group were given intraventricular injection of siNgR and transfection reagents ( 10μl ); the rats of NS group were given intraventricular injection of NS and transfection reagents (10μl);and the rats of HIBD group had no intervention. In addition to these three groups, there is another group, sham-operated group ( n=10 ) . The rats of sham-operated group were sham-operated ( common carotid artery was isolated but not ligated) and did not receive hypoxia-ischemia processing and intraventricular injection. Utilize water maze experiment to analyze the rats' escape time. The levels of NgR and GAP-43 protein in rats' brains were measured by immunohistochemistry and image analysis. Results RT-PCR gel electrophoresis results showed that the NgR cDNA stripe of siNgR group was not obvious, but the stripe of NS group was clear. At the same time, the GAPHD cDNA bands of the above two groups were both clear. There were more NgR positive immune reaction products ( brown particles) in NS group than in siNgR group. The number of GAP-43 positive cells by immunohistochemistry in sham-operated group, HIBD group, NS group and siNgR group was (33. 24±1. 32), (20. 14±1. 24), (18. 73±1. 41) and (28. 06±1. 78), respectively. The number of sham-operated group and siNgR group was greater than HIBD group and NS group ( P＜0. 05 ) . There was no statistical significant difference for the number of GAP-43 positive cells between sham-operated group and siNgR group ( P＞0. 05 ) . Water maze experiment results showed that the newborn rats ' average escape time ( s ) of HIBD group ( 58. 1 ± 10. 3,47. 2±10. 1, 42. 5±7. 6) was obviously longer than sham-operated group (34. 2±5. 6, 25. 7±6. 2, 21. 2±8. 1), and the difference was statistically significant (P＜0. 05). However, the average escape time of siNgR group (37. 5±9. 8, 29. 1±9. 8, 27. 2±9. 3) was obviously shorter than HIBD group and NS group (60. 7±5. 2, 49. 1±9. 9, 45. 3±9. 3), (P＜0. 05). Conclusions Chemically synthesized specific siRNA had the potential to interference the expression of NgR in the brain of newborn rats, and to a certain extent, could promote the nerve regeneration and neural functional recovery of rats.

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The influence of p53 protein expression on kidney development in rats with intrauterine growth restriction

Yong JI ; Jianfang GE ; Rongwei MA ; Jianhong YAO

Chinese Journal of Neonatology.2015;30(3):218-223. doi:10.3969/j.issn.1673-6710.2015.03.019

Objective To study the effect of renal p53 protein expression on kidney development in rats with intrauterine growth restriction ( IUGR) . Methods Pregnant rats were randomly assigned into normal group, IUGR group and L-Arg treated group. Normal group were fed with normal diet (21％protein). IUGR group and L-Arg treated group were fed with low-protein diet (10％ protein). During lactation, maternal rats in the three groups were all fed with normal diet. Maternal rats in L-Arg treated group were given additional special drinking water ( L-Arg:200 mg/kg) , while maternal rats in normal and IUGR group were given normal drinking water. Offspring weaned after 21 d and had free access to normal rodent diet and water. When the pups grew up to 2 m, the number of glomeruli was counted using acid digestion method, the proliferation and apoptosis of glomerular and tubular cells were studied using TUNEL and Ki-67 immunohistochemistry, and the ultrastructure of epithelial cells was determined by electron microscopy. At 7 d, 21 d, 2 m and 3 m, p53 protein expression in renal tissue was measured by Western blot, respectively. Results At 2 m, the number of glomerulus ( right kidney) in IUGR group was significantly lower than normal group [(23 647±541) vs. (27 689±492), P＜0. 01]; the index of renal cell apoptosis in IUGR group was higher than normal group [(21. 9 ± 2. 0) vs. (16. 7 ± 2. 5), P＜0. 05];however, IUGR group and normal group had no statistically significant difference (P＞0. 05). The proliferation of mesangial cells was found in IUGR and L-Arg treated group, but not in normal group. And the extent of proliferation in L-Arg treated group was lesser than IUGR group. A reduction of foot process and partial fusion of foot process could be observed in IUGR group and L-Arg treated group while the foot process morphology in normal group was normal. At 2 m and 3 m, p53 protein level in IUGR group and L-Arg treated group were higher than normal group [ 2 m: ( 0. 28 ± 0. 03 ) and ( 0. 21 ± 0. 01 ) vs. (0. 10±0. 02);3 m:(0. 39±0. 04) and (0. 26±0. 02) vs. (0. 17±0. 03);P＜0. 01], while IUGR group higher than L-Arg treated group ( P＜0. 05 ) . Conclusions The kidney of IUGR rats showed reduced glomerular number, increased renal cell apoptosis, enhanced p53 protein expression, increased proliferation of glomerular mesangial cells, decreased foot process, and partial fusion of foot process. And L-Arg could to some extent improve the organizational structure of the kidney in IUGR rats.

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A study on the treatment of infections in neonatal surgical patients

Yu WANG ; Xianqing JIN ; Li XIANG ; Wei LIU

Chinese Journal of Neonatology.2015;30(3):200-202. doi:10.3969/j.issn.1673-6710.2015.03.013

Objective To study the feature of neonatal infections and characteristics of antibiotic treatment in a tertiary children ' s hospital. Methods Clinical data including incidence of infection, primary disease,species of bacteria, complication and antibiotic utilization in hospitalized patients from Jan. 2010 to Dec. 2012 were retrospectively reviewed using their medical records. Results Among 1826 patients admitted to neonetal surgery ward, 542 infants ( 29. 7％) were with infection. The incidence of antibiotic resistance was 23. 51％. The top five infectious diseases were:perianal abscess, necrotizing enterocolitis, colicitis, omphalitis and subcutaneous gangrene. 12 cases of multi-resistant infection were cured by non-restricted antibiotics. 109 were cured by restricted antibiotics. And other 7 were cured by special antibiotics. No death nor multi-resistant nosocomial infection were found. Risk factors including multi-site infection, premature or low birth weight infants, liver, kidney or heart dysfunction,fever lasting more than 3 days after antibiotic therapy, septic shock, sepsis, digestive tract perforation and peritonitis,were vital in choosing specific antibiotics. Conclusions Infection is one of the most common diseases in neonatal surgery ward, with major pathogens sensitive to antibiotics. The clinical characteristics and drug sensitive test are conductive to the reasonable use of antibiotics. Special antibiotics can be used directly in patients with risk factors Clinical doses of antibiotics in neonates depend on the monitoring of drug concentration.

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The value of serum albumin level for the prognosis of late-preterm infants infection

Chunyan YANG ; Baoyun LI ; Ping XU ; Yujun YANG ; Qiaozhi YANG

Chinese Journal of Neonatology.2015;30(3):196-199. doi:10.3969/j.issn.1673-6710.2015.03.012

Objective To study the value of serum albumin ( ALB ) level for the prognosis of late-preterm infants infection. Methods Late-preterm infants admitted to the neonatal intensive care unit ( NICU) from July 2012 to July 2013 were recruited and their clinical data retrospectively reviewed, including the laboratory examination results, neonatal critical illness scores ( NCIS ) , perinatal complications and prognosis. The infants were assigned into three groups based on ALB levels (＞30 g/L, 25-30 g/L, ＜25 g/L). Results A total of 257 cases were recruited and 122 cases (47. 4％) had ALB levels ＜25 g/L. 32 had neonatal sepsis ( sepsis group) , 190 neonatal infection ( infection group) and 35 without infection ( no-infection group ) . The incidences of hypoalbuminemia among these groups were 84. 4％, 50. 0％ and 28. 6％, with the mortality rate 15. 6％, 0. 5％ and 0％. The incidence of hypoalbuminemia and mortality rate in sepsis group were significantly higher than the other groups ( P＜0. 05 ) , and no statistically significant differences between infection group and no-infection group ( P＜0. 05). The ALB level in survived infants [(29. 6±7. 5)g/L] was statistically higher than the deceased ones [(20. 4±6. 9)g/L](P＜0. 05). The incidence of critically ill newborns was 65. 5％ in ALB ＜25 g/L group, significantly higher than the other groups (P＜0. 05). 26. 2％ in ALB ＜25 g/L group had more than 4 organs injuries, significantly higher than ALB ＞30 g/L group ( P＜0. 05 ) . Conclusions Hypoalbuminemia is common among neonates with sepsis. The ALB level had predictive value for the prognosis of neonatal infection.

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Relationship between genetic mutations and therapeutic effect of glibenclamide in permanent neonatal diabetes

Huimin HAO ; Dongxia FU ; Ting HUANG ; Haiyan WEI

Chinese Journal of Neonatology.2015;30(3):192-194. doi:10.3969/j.issn.1673-6710.2015.03.010

Objective To study the diagnostic significance and clinical value of genetic analysis in children with neonatal diabetes. Methods Gene mutation analysis was performed in four patients from Zhengzhou children ' s hospital with diagnosis of with neonatal diabetes. Therapeutic effect of glibenclamide in patients with or without gene mutation was compared. Results KCNJ11 gene mutation was found in two patients with neonatal diabetes. Glibenclamide was found only effective for blood glucose control in patients with KCNJ11 mutation. Therefore, Insulin remains the best therapeutic choice in patients without the genetic mutation. Conclusions Genetic mutation status may be useful in choosing treatment options of neonatal diabetic patients, therefore, should be performed in all children with neonatal diabetes.

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Correlation analysis between early changes in biochemical markers of bone metabolism and early nutrition of preterm infants

Huabao PENG ; Wen XIA ; Ruokun TAN ; Liwei TANG ; Ning TAN

Chinese Journal of Neonatology.2015;30(3):188-191. doi:10.3969/j.issn.1673-6710.2015.03.009

Objective To explore the changes of bone metabolic markers during early stage of preterm infants, as well as the relationship with their nutrition status. Methods Preterm infants with gestational age 30-35 weeks admitted to our Hospital were collected from November 2012 to April 2013. Venous blood samples obtained within 24 hours after birth and between 8:00-9:00 AM two weeks after birth were used to determine the Serum β-CTx, osteocalcin ( OC) and propeptide of type I procollagen (PINP) levels by Electro-Chemiluminescence. Analysis of changes of these bone metabolic markers and their relationship with early stage nutrition related indicators were also performed. Results A total of 60 premature infants were collected. There was no significant correlation among serum β-CTx, OC and PINP within 24 hours after birth ( r=0. 170, P＞0. 05 ) . The Serum β-CTx within 24 hours after birth was negatively correlated with gestational age (r= -0. 603, P＜0. 05), whereas the serum OC within 24 hours after birth was positively correlated with gestational age ( r=0. 581, P＜0. 05 ) . However, PINP wasn′t correlated with gestational age significantly (r=0. 134,P＞0. 05). Serumβ-CTx, OC and PINP at 2 weeks after birth increased significantly from the baseline level detected within 24 hours after birth ( P＜0. 05 ) .Δβ-CTx was positively correlated with ΔOC (r=0. 600,P＜0. 05). There was no significant correlation between ΔPINP and Δβ-CTx (r=0. 045,P＞0. 05), as well as ΔOC and ΔPINP (r=0. 110,P＞0. 05).ΔOC was positively correlated with average daily calorie ( P＜0. 05 ) and average daily P/E ( P＜0. 05 ) , negatively correlated with cumulative loss of caloric ( P＜0. 05 ) . There was no significant correlation between Δβ-CTx or ΔPINP with nutrition related indicators of this study. Conclusions Serum OC within 24 hours after birth of preterm infants and their gestational age are positively correlated, while β-CTx detected at the same time and gestational age are negatively correlated. Vigorous metabolism of premature bone occurs during the first two weeks after birth, as the serum β-CTx, OC and PINP levels increased significantly. We suggest that reasonable calorie intake and appropriate protein calorie ratio at early stage after birth is very important for the development of bone of preterm infants.

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A study on the effects of four major neonatal diseases on thyroid function

Xiangming ZHONG ; Minhong LI ; Xun CHEN ; Zhiqing DENG ; Zhihong. LUO

Chinese Journal of Neonatology.2015;30(3):183-187. doi:10.3969/j.issn.1673-6710.2015.03.008

Objective To study the effects of preterm birth, infection, pathological jaundice and asphyxia on neonatal thyroid function. Methods From April 2012 to April 2014,neonates within 7 days after birth admitted to the neonatal department of our hospital were recruited. The neonates with one or more of the 4 aforementioned conditions were assigned to the observation group; the full-term newborns without any diseases to the control group. 2 ml of venous blood from each patient was collected 5-7 days after birth to determine the levels of serum free iodine thyroid original glycine ( FT3 ) , free thyroxine ( FT4 ) and thyroid stimulating hormone ( TSH ) . The thyroid function of newborns in the observation group were re-examined after recovery, and compared among different groups and conditions. Results A total of 220 newborns were recruited in the observation group and 34 in the control group. FT3 in the observation group was significantly lower than the control group ( P＜0. 05 ) . As for FT4 and TSH, no statistically significant differences existed between the two groups ( p＞0. 05 ) . FT3 and FT4 in neonates with severe infection were significantly lower than mild infection ( P＜0. 05 ) . FT3 in the preterm group with gestational age ( GA ) ≤32 weeks was significantly lower than GA＞32 weeks, and FT3 in the jaundice group, serum bilirubin level ≥ 342 mol/L group significantly lower than ＜342 mol/L group (P＜0. 05) . In the observation group, the re-examined FT3 concentration was significantly higher (P＜0. 05) than the result tested 5-7 days after birth while FT4 and TSH didn′t show significant differences. And no significant differences existed between the observation group and the control group in re-examined thyroid functions ( P＞0. 05 ) . The multivariate analysis showed that GA, pathological jaundice, infection and asphyxia were associated with FT3 concentration 5-7 days after birth (P＜0. 05), while the delivery method and gender not (P＞0. 05). Conclusions The main effect of preterm birth, infection, pathological jaundice and asphyxia on thyroid function was to decrease FT3 temporarily. Severe infection might decrease FT4 temporarily. However, the thyroid function would return to normal with recovery.

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A clinical study of the brain natriuretic peptide in the diagnosis of myocardial injury after neonatal asphyxia

Rui ZHU ; Zhenhong NIE ; Dongchi ZHAO

Chinese Journal of Neonatology.2015;30(3):178-182. doi:10.3969/j.issn.1673-6710.2015.03.007

Objective To investigate the relationship between serum plasma brain natriuretic peptide ( BNP ) levels and myocardial injury in neoborns after asphyxia. Methods Neoborns who were admitted to Department of neonatology, Zhongnan Hospital, Wuhan University from December 2012 to December 2013 within 3 days after birth were considered. According to the number organized in chronological order every other case, newborns with neonatal asphyxia were assigned to observation group. The observation group were further divided into myocardial injury subgroup and non-cardiac injury subgroup according to the diagnostic criteria of myocardial injury. Newborns without neonatal asphyxia or neonatal cardiovascular diseases were assigned to control group. Exclusion criteria for control group were electrolyte disturbance, liver and kidney dysfunction. Blood sample was drawn from patients within 2 hours of admission to hospital and again on day 14. Serum BNP , creatine kinase isoenzyme ( CK-MB) , serum sodium and calcium were detected for further analysis. Results In 107 cases with neonatal asphyxia, 77 infants who had complete clinical records were selected as observation group, of which 36 met the diagnostic criteria of myocardial injury and assigned to myocardial injury subgroup. Non-cardiac injury subgroup consisted of the rest 41 cases in observation group. Twenty-two cases were enrolled to control group. Within 2 hours after admission, the serum BNP level of myocardial injury subgroup were significantly higher than those of the non-cardiac injury subgroup and the control group ( 2. 35 ± 0. 44 , 2. 12±0. 64, 1. 88±0. 27, log transformed, respectively, P＜0. 05). The BNP level of non-cardiac injury subgroup were also significantly higher than those of the control group. Serum BNP and CK-MB levels of observation group were positively correlated (r=0. 212,P=0. 030). After treatment, serum BNP level of myocardial injury subgroup at 14 days after admission decreased significantly, compared to the level at 2 hours within admission (P＜0. 05). When the cutoff value for infants with myocardial injury was 108. 05 pg/mL, the area under the ROC curve was 0. 753, with a sensitivity of 75. 0％ and a specificity of 64. 5％, positive predictive value was 56. 4％ and negative predictive value was 72. 3％. Conclusions Serum BNP level can reflect myocardial injury in neonates with asphyxia and can guide clinical treatment.

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The roles of hypoxia-inducible factor-1 alpha and vascular endothelial growth factor in the pathogenesis of hypoxia-induced pulmonary hypertension in neonatal rats

Yili MA ; Le WANG ; Mingxia LI

Chinese Journal of Neonatology.2017;32(1):64-68. doi:10.3969/j.issn.2096-2932.2017.01.016

Objective To studytherolesof hypoxia-inducible factor-lalpha( HIF-1α )and vascular endothelial growth factor ( VEGF) in the pathogenesis of hypoxia-induced pulmonary hypertension ( HPH) in neonatal rats.Methods Wistar neonatal rats were assigned into HPH group and the control group using random number table method. Neonatal rats in HPH group were exposed to hypoxia according to HPH model. On day 3 , 7 ,14 and 210f hypoxia,the mean pulmonary artery pressure ( mPAP) , the level of mRNA and protein expression of HIF-1α and VEGF in lung tissue were examined using RT-PCR and Western blot methods respectively. The correlation of HIF-1,VEGF and mPAP were also analyzed. Results The mPAP ( mmHg)in HPH group on day 3 ,7 ,14 and 210f hypoxia were all higher than the control up[8.5±1.5)vs.5.2±1.0),(12.1±2.1)vs.9.6±0.8),(12.9±2.0)vs.(9.1±0.8),(21.0±2. 3) vs. (11. 2 + 1. 6) ,P < 0. 05]. 0n day 3, 7 and 140f hypoxia,the mRNA of HIF-1α in lung tissue of HPH group were significantly higher than the control group ( P <0. 05 ) . On day 70f hypoxia, the HIF-1αprotein in lung tissue of HPH group was significantly higher than the control group(P <0. 05 ) . On day 7 ,14 and 210f hypoxia,the mRNA and protein of VEGF in lung tissue of HPH group were also significantly higher than the control group( P <0. 05 ). Correlation analysis showed that HIF-1α protein were positively correlated with mPAP on day 3 ,7 . 14 and 210f hypoxia in HPH group ( r = 0. 504 .P = 0. 002) , and VEGF protein were positively correlated with mPAP in HPH group on day 7 , 14 and 210f hypoxia( r = 0. 782, P < 0. 001) .Conclusion Both HIF-1α and VEGF play roles in the occurrence and development of HPH in neonatal rats.

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Neurobehavior and motor development changes in the 3-day-old rats with ischemic brain injury

Lan HU ; Chao CHEN

Chinese Journal of Neonatology.2017;32(1):59-63. doi:10.3969/j.issn.2096-2932.2017.01.015

Object To investigate the pathological changes,physiological condition,neurobehavior and motor development of the 3-day-old rats with ischemic brain injury.Methods Ninety six 3-day-old male Sprague-Dawley rats were randomly assigned to experimental and control groups.Occlusion of both carotid arteries was performed in the experimental group,while the rats in the control group only received skin incisions without carotid ligation.Physical examinations and neurobehavioral development of the rats were recorded daily from the first day after operation until 3 weeks after birth,including weight,eye opening, incisor eruption,ear unfolding,righting reflex,negative geotaxis,limb placing and grasping reflex. Specimens of the brain tissue were obtained in 24 hours after operation and 3 weeks after birth for the hematoxylin-eosin staining and immunohistochemistry staining to investigate the pathological changes.All the reaults were compared between the 2 groups.Results Compared with the control group,rats in the experimental group were found growth retardation,suppression of primitive reflexes and impaired motor abilities (P < 0.05 ).The brain tissue obtained from the rats after operation showed white matter rarefaction,liquefaction and microglia hyperplasia with Hematoxylin-eosin staining and myelin formation disorder with immunohistochemistry staining.Conclusions Ischemic brain injury of the 3-day-old rats could result the mental retardation,neurobehavioral and motor development disorder because of the white matter injury.

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