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A study on changes of brain tissue inflammatory cytokines in neonatal rats with periventricular leukomalacia

Jian LI ; Ying OUYANG ; Ke HUANG ; Hong DONG ; Qin WANG

Chinese Journal of Neonatology.2017;32(2):150-154. doi:10.3760/cma.j.issn.2096-2932.2017.02.017

Objective To study inflammatory cytokines changes in brain tissue of neonatal rats with periventricular leukomalacia (PVL).Method A total of 80 neonatal SD rats (P3) were randomly assigned into 2 equal groups,sham-operated group and PVL group.Rats in each group were further assigned into four subgroups (12,24,48,72 h),with 10 rats in each subgroup.The hypoxic-ischemic PVL modal were established following the procedure:first,isolation and ligation of left common carotid artery,and then exposed to 8％ O2 and 92％ N2 for 2.5 h.The sham-operated rats were processed with isolation of left common carotid artery only.Rats of the four subgroups were sacrificed at 12,24,48 h and 72 h respectively,then the brains were rapidly removed in corresponding time.Pathological changes of brain tissues were observed using HE stain.The mRNA expression levels of tumor necrosis factor α(TNF-α)and inter leukin-1β (IL-1β) were assessed using real-time quantitative PCR assays,the protein levels of TNF-α and IL-1β were detected using enzyme linked immunosorbent assay method.Result The brains tissues of rats in PVL group showed remarkably hyperemia and edema,with left ventricle enlargement.Periventricular white matter structure was disintegrated comparing with sham-operated group.The expression of TNF-α and IL-1 β mRNA in PVL group increased significantly,reaching peak by 24 h and then gradually decreased 72 h after the procedure.The mRNA levels of TNF-c and IL-1 β were significantly different between each two time points of 12,24 h and 48 h in PVL group (P ＜0.05).However,there was no differences between 72 h and 48 h within PVL and sham-operated group group(P ＞0.05).In PVL group,the protein expression trends of TNF-α and IL-1 β were similar to mRNA expression trends.Moreover,the protein levels were significantly different between each two time points of TNF-α and IL-1 β,respectively (P ＜ 0.01).The protein expression levels of TNF-α were different at each time point between PVL group and sham-operated group[(189.2 ± 20.4) pg/ml vs.(131.4 ±5.2) pg/ml at 12 h,(213.8 ± 16.7) pg/ml vs.(127.7 ±7.4) pg/ml at 24 h,(181.7 ± 15.0) pg/ml vs.(126.3 ± 6.0) pg/ml at 48 h,(159.6 ± 25.3) pg/ml vs.(131.4 ± 6.0) pg/ml at 72 h;P ＜0.01].The protein levels of IL-1β were different between the two groups only at 24 h and 48 h.[(121.8 ±30.0) pg/ml vs.(67.4 ± 13.7) pg/ml,(83.3 ± 15.7) pg/ml vs.(65.3 ± 14.9) pg/ml;P ＜0.05].In sham-operated group,no differences of TNF-α and IL-1 β protein levels were found between any different time points (P ＞ 0.05).Conclusion Inflammatory cytokines such as TNF-α and IL-1β are involved in ischemic-hypoxia induced PVL.Dynamic detection of inflammatory factors is expected to be an important method of early diagnosis,assessment of treatment efficacy and prognosis of PVL.

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Neuroprotective effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats

Chao HUANG ; Yinxiang YANG ; Zhaoyan WANG ; Qian WANG ; Jingguo CHEN ; Zuo LUAN

Chinese Journal of Neonatology.2017;32(2):144-149. doi:10.3760/cma.j.issn.2096-2932.2017.02.016

Objective To explore the effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats and to compare the difference of therapeutive effect between the cerebellum medulla oblongata pool injection and the jugular vein injection.Method A total of 73 three-day-old SD rats were chosen to establish the model of white mater injury.After 24 hours,the 73 rats were randomized into the experimental group (n =46) and the control group (n =27).Then the experimental group was reclassified into ventricular group (n =23) and intravenous group (n =23).In the ventricular group,the paracrine extracts of human adipose stem cells was injected locally into the cerebellum medulla oblongata pool injection,while the extracts was injected into the jugular vein in the intravenous group.The control group was reclassified ventricular control group (n =15) and intravenous control group (n =12),and equivoluminal saline was injected the same way as the experimental group.Frozen sections of the brain tissue from 3 rats of each experimental group one day after injection were stained with fluorescein-conjugated streptavidin to study the distribution of the extracts.The brain tissue of 3 rats from each subgroup 3 days after injection were stained with hematoxylin eosin (HE) to observe the pathomorphological changes.While 7 days later,myelin basic protein (MBP) of white matter which was obtained from 7 rats of each group was detected by immunofluorescence staining.28 days after injection,the remaining rats were assessed by neurobehavior tests.For the rats that died during the experiment,the same number of the rats would be substituted in this study.Result The paracrine extracts were found to transfer to the brain lesion area,and the amount of the extracts was more in the ventricular group.The results of the HE staining showed that the white matter injury was more severe in the ventricular control group,and extensive area of infarction were found in this group.White matter injury was mild in the experimental group,and the structure of the corpus callosum was more complete in the ventricular group.MBP semi-quantitative scores of the ventricular group (0.7 ± 0.3) and intravenous group (1.7 ± 0.3) were lower than those of ventricular control group (3.4 ± 0.4)and intravenous control group(3.3 ±0.3).And the MBP scores of ventricular group was significantly lower than that of intravenous group (P ＜ 0.05).The scores of the neurobehavioral tests of the experimental group were significantly higher than those of the control group,while the scores of the ventricular group were significantly higher than those of the intravenous group (P ＜ 0.05).Conclusion The paracrine extracts derived from adipose stem cells could improve the prognosis of white matter injury,and cerebellum medulla oblongata pool injection showed better curative effect than the jugular vein injection.

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The effects of delayed cord clamping in preterm infants: a meta-analysis

Yiyu CHEN ; Jin ZHOU ; Yuwei HE

Chinese Journal of Neonatology.2017;32(2):137-143. doi:10.3760/cma.j.issn.2096-2932.2017.02.015

Objective To study the short-term effects of delayed cord clamping (DCC) in preterm infants.Method A thorough search was conducted on medical databases including Cochrane Library,PubMed,Ovid,Medline,VIP citation databases,Wanfang database and CNKI.Randomized control trials (RCTs) of DCC in preterm infants were retrieved from medical literature published during January 1,2000 to January 1,2016.DCC group had cord clamping 30 ～60 s after birth,and immediate cord clamping (ICC) group had cord clamping within 30 s after birth.Methodological quality was evaluated using Cochrane Handbook for systematic reviews and RevMan 5.1 software.Meta-analysis was performed using RevMan 5.1 software.Result Seventeen RCTs were included.Meta-analysis showed that:the blood pressure within 4 hours after birth (WMD =2.49,95％ CI 0.74 ～ 4.24),the hemoglobin concentration (WMD =15.92,95 ％ CI 6.37 ～ 25.47) and the hematocrit (WMD =4.84,95 ％ CI 3.47 ～ 6.22) within 24 hours after birth in the DCC group were higher than the ICC group,P ＜0.05;the risk of anemia (RR =0.62,95％ CI 0.47 ～ 0.81),intraventricular hemorrhage (RR =0.64,95 ％ CI 0.45 ～ 0.91) and mortality (RR =0.42,95％ CI 0.20 ～0.86) in the DCC group were lower than the ICC group,P ＜0.05;there were no statistically significant differences between the two groups in peak of serum bilirubin,phototherapy duration,rate of phototherapy treatment and blood transfusion,the incidence of hyperbilirubinemia and polycythemia (P ＞ 0.05).Conclusion DCC is safe and feasible for premature infants,and can improve the outcome of premature infants.

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Case report of congenital glucose-galactose malabsorption in China and literature review

Li TAO ; Ling WANG ; Xiaowen CHEN ; Ning ZHAO ; Lanlan GENG ; Jianfeng LIN ; Yan WEN ; Wei ZHOU

Chinese Journal of Neonatology.2017;32(2):123-127. doi:10.3760/cma.j.issn.2096-2932.2017.02.011

Objective To study the clinical features of congenital glucose-galactose malabsorption (CGGM),and to improve the understanding of CGGM.Method Clinical manifestations and treatment process of one patient with CGGM in our hospital were retrospectively analyzed.From 1966 to 2016 May,Chinese medical database and PUBMED were searched using Malabsorption syndrome,dehydration,hypernatremia , diarrhea , newborn , carbohydrate metabolism ,andglucose/galactose malabsorption as key words.The clinical features of CGGM reported in literatures were summarized.Result The patient in our hospital was a full-term female infant naturally delivered.The onset of the disease was on the 9th day after birth,and the clinical manifestations included severe diarrhea,severe dehydration,hypernatremia,metabolic acidosis and malnutrition.After intravenous infusion and symptomatic treatment,dehydration,hypernatremia and metabolic acidosis were corrected.However,there was no improvement of diarrhea characterized with watery and acidic stools,and neither was weight gain.Glucose loading test was negative,and fructose loading test was positive.Diarrhea was improved markedly using diagnostic carbohydrate-free formula,so CGGM was diagnosed clinically.SLC5A1 homozygous IVS7-2 A ＞ G mutation was detected which confirmed the diagnosis of CGGM.With carbohydrate-free formula feeding,the body weight of the infant was increased.Followed up for 2 months now,her body length and body weight were at P25 and P22 on growth curve respectively,and no obvious neurological sequela was observed.Our literature review revealed 7 reports including 48 cases of CGGM children.Literature review showed that:most children with CGGM (79.2％) had the onset within 7 days of life;main clinical features included diarrhea (100％),dehydration (100％),and malnutrition (54.2％);22.9％ of patients with carbohydrate-free formula and 27.1％ with fructose matrix formula were fed well;no death was detected,77.1％ had normal weight gain,and 91.7％ had normal development of the nervous system.Conclusion CGGM is rare.The symptoms include severe watery and acidic stools with onset during neonatal period.CGGM is associated with severe complications such as hypertonic dehydration and hypernatremia.The diagnosis is established based upon typical clinical manifestations,sugar loading test and SLC5A1 gene detection.Carbohydrate-free formula feeding is effective.

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The application of array comparative genomic hybridization for the detection of chromosomal defects in newborns

Min JIANG ; Yan ZHONG ; Yujie QI ; Yajuan WANG ; Caiyun YANG

Chinese Journal of Neonatology.2017;32(2):119-122. doi:10.3760/cma.j.issn.2096-2932.2017.02.010

Objective To study the application of the array comparative genomic hybridization (Array-CGH) for the detection of chromosomal disorders in newborns.Method The Array-CGH technique was used to analyze the whole genome of the patients who were suspected of chromosomal disease in neonatal ward of our hospital from January to December in 2014,and further verification in genomic unbalanced ectopia was carried out by FISH (fluorescence in situ hybridization,FISH).Result Among 514 patients,104 were found carrying chromosomal abnormalities with a detection rate of 20.2％.The most common chromosomal disease is the Down syndrome syndrome (24 cases),followed by the chubby Willy and Angel syndrome(17 cases),while the Wolf-Hirschhorn syndrome in 5 cases,Williams syndrome in 5 cases and the Criduchat syndrone in 5 cases.The results of FISH were consistent with Array-CGH.Conclusion The technique of Array-CGH can be used to scan the whole genome of children with unknown disease.As a high-throughput and rapid research method,this technique has important clinical significance in the screening of chromosomal diseases.

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Relationship between early-onset neonatal sepsis and the level of interleukin-6, C-reactive protein in umbilical cord serum of the newborns with premature rupture of membrane

Changyi YANG ; Baoquan ZHANG ; Hanqiang CHEN

Chinese Journal of Neonatology.2017;32(2):110-114. doi:10.3760/cma.j.issn.2096-2932.2017.02.008

Objective To study the levels of interleukin-6 (IL-6) and C-reactive protein(CRP) in umbilical cord serum of the newborns with premature rupture of membrane(PROM)and to explore the value of IL-6 and CRP in the diagnosis of early onset neonatal sepsis (EONS).Method A total of 187 term newborns with PROM ＞ 12 h who were born normal vaginally in our Hospital from April 2015 to December 2015 were enrolled in this study as the PROM group and another 50 term infants without PROM and infection as the control group.The levels of IL-6 and CRP in umbilical cord serum were quantified by ELISA,the results of which were compared between groups.Receiver operating characteristics (ROC) curves were drawn to find out the cut-off value of IL-6 and CRP for the diagnosis of EONS.Result The levels of IL-6 and CRP in umbilical cord serum in the PROM group were significantly higher than those in the control group [IL-6 20.3 (9.5,35.8) pg/ml vs.9.3 (6.9,27.5) pg/ml,CRP 0.42 (0.25,0.78) mg/L vs.0.33 (0.18,0.45) mg/L,P ＜ 0.05].The levels of IL-6 and CRP in the newborns whose mother had chorioamnionitis were significantly higher than those in the newborns whose mother was without chorioamnionitis [IL-6 62.5 (35.2,92.7) pg/ml vs.10.8 (9.3,33.4) pg/ml,CRP 0.86 (0.44,1.95) mg/L vs.0.35 (0.20,0.62) mg/L,P ＜0.05].The levels of IL-6 and CRP in the infants with PROM≥18 h was significantly higher than those in the infants with PROM ＜ 18 h [IL-6 32.1 (9.9,42.2) pg/ml vs.10.7 (9.2,32.6) pg/ml,CRP 0.44(0.29,0.86) mg/L vs.0.35 (0.23,0.61) mg/L,P ＜ 0.05].The levels of IL-6 and CRP in the neonates with EONS was significantly higher than those in the neonates without EONS [IL-6 92.0 (58.3,161.0) pg/ml vs.20.0(9.4,35.2)pg/ml,CRP 1.94(0.47,2.73) mg/L vs.0.38(0.24,0.67) mg/L,P ＜ 0.05].ROC curve analysis showed that the cut-off value of IL-6 and CRP for the diagnosis of EONS were 81.lpg/ml (sensitivity 76.5％,specificity 90.6％) and 1.88mg/L (sensitivity 64.7％,specificity 89.4％).With the combination of IL-6 and CRP levels,the sensitivity was 88.2％ and the specificity was 84.1％ for the diagnosis of EONS.Conclusion To measure the IL-6 and CRP levels in umbilical cord serum is helpful for the early diagnosis of EONS,and the combined detection of the 2 items may improve the sensitivity of diagnosis.

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A study on the interleukin-10 receptor gene mutations and neonatal very early onset inflammatory bowel disease in 5 infants

Yi JIANG ; Donghui CHEN ; Lili LIU ; Xinlin HOU ; Zezhong TANG ; Congle ZHOU

Chinese Journal of Neonatology.2017;32(2):105-109. doi:10.3760/cma.j.issn.2096-2932.2017.02.007

Objective To study the clinical features,diagnosis and treatment of neonatal very early onset inflammatory bowel disease(VEO-IBD) to improve the diagnosis and treatment of the disease.Method From Jan 2013 to Dec 2015,five infants with VEO-IBD admitted to Peking University First Hospital were reviewed and analyzed.Their clinical data included general condition,clinical symptoms,laboratory tests,autoimmune antibodies (ANCA,dsDNA,ANA and ENA),colonoscopy,pathological results and therapeutic response.Interleukin-10 receptor A (IL-10RA) gene was examined in all patients.All the patients were followed up for more than 1 year.Result Three of the five patients had a family history.Persistent diarrhea was the most common presenting symptom.One of them received surgery because of intestinal necrosis and developed typical symptoms of IBD half a year later.Bloody stool or positive fecal occult blood test were found in all infants.Fever,anemia,oral ulcer,perianal lesions and malnutrition were common concomitant symptoms.Most of the patients had elevated WBC,CRP and ESR,and 4 of them had positive autoimmune antibodies.Colonoscopy showed multiple ulcers affecting the colon.Intestinal biopsies revealed acute and chronic inflammation.4 of patients were found to have cryptitis and crypt abscesses.Gene sequencing revealed IL-10RAgene mutation in all five patients,including 1 case with homozygous mutation and 4 heterozygous mutations.4 patients received steroid and mesalazine therapy and only 1 patient's symptoms were controlled.However,the colonoscopy result was still abnormal in this patient.4 patients had poor response to further infliximab and (or) thalidomide therapy.1 of them received surgery because of intestinal obstruction at 2-year-old.Conclusion Neonatal VEO-IBD was associated with IL-10RA gene mutation.The patients had severe symptoms and poor response to conventional medications.The effects of biological agents and thalidomide were still not sure.

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Effect of three different modes of non-invasive positive pressure ventilation on cardiac indices of premature infant with respiratory distress syndrome: a randomized control study

Honglin LEI ; Xiangyu GAO ; Di HUANG ; Dandan ZHAO ; Bo YANG ; Yi REN ; Niannian TONG

Chinese Journal of Neonatology.2017;32(2):100-104. doi:10.3760/cma.j.issn.2096-2932.2017.02.006

Objective To study the effect of three different modes of non-invasive positive pressure ventilation on cardiac indices of premature infants with respiratory distress syndrome (RDS).Method From January 2014 to October 2015,preterm infants who had RDS received intubation-pulmonary surfactantextubation in the neonatal intensive care unit of the Hospital were randomly assigned (by random number table) to three groups based on the primary mode of ventilation:nasal continuous positive airway pressure (NCPAP),bi-level positive airway pressure (BiPAP),and synchronized bi-level positive airway pressure (SBiPAP).The mean airway pressure (MAP) were about 6 cmH2O in the three groups.The level of plasma B-type natriuretic peptide (BNP),cardiac troponin Ⅰ (cTnI),and correct QT intervals dispersion (QTcd) were monitored before and 42-54 h after non-invasive ventilation.Result There were 173 cases in our study,59 of which in NCPAP group,56 in BiPAP group,and 58 in SBiPAP group.The plasma BNP level at 42-54 h after non-invasive ventilation in the three groups were all higher than that before non-invasive ventilation [NCPAP group:(247.9 ± 137.9) ng/L vs.(182.5 ± 1 10.7) ng/L,P =0.007;BiPAP group:(258.5 ± 131.2) ng/L vs.(182.6 ± 105.0) ng/L,P ＜ 0.001;and SBiPAP group:(260.9 ± 159.7) ng/L vs.(177.5 ± 101.5) ng/L,P =0.002].After 42-54 h non-invasive ventilation,there were no significant changes of plasma cTnI level and QTcd in all the three groups (all P ＞ 0.05).The level of plasma BNP,cTnI,and QTcd among the three groups before and after 42-54 h non-invasive ventilation all showed no significant differences statistically (all P ＞ 0.05).Conclusion Longer duration (42-54 h) of non-invasive positive pressure ventilation (MAP:6 cmH2O) in preterm infants with RDS may lead to increased plasma BNP level,and may affect their cardiac function.However,it may not lead to serious myocardial damage and abnormality of ventricular repolarization.There were no significant differences in cardiac indices of premature infant with RDS among NCPAP,BiPAP,and SBiPAP group with the same MAP.

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The clinical characteristics of neonatal sepsis caused by different pathogens

Xiaoying CHEN ; Lihua QIU ; Qiannan JIANG ; Lisheng ZHANG ; Ke YUAN

Chinese Journal of Neonatology.2017;32(2):115-118. doi:10.3760/cma.j.issn.2096-2932.2017.02.009

Objective To study the characteristics of neonatal sepsis caused by gram positive (G +) bacteria,gram negative (G+) bacteria and fungi.Method Clinical data of 202 neonates with sepsis hospitalized from Jan.2012 to May.2015 were studied.According to the different pathogens,202 neonates were divided into gram positive bacteria group,gram negative bacteria group and fungi group.The general information,clinical manifestation,laboratory examination and treatment outcome of the three groups were analysed with Chi square analysis,LSD,Fisher exact probability tests.Result A total of 202 cases of neonatal sepsis were recruited.The detection rate of gram positive bacteria,gram negative bacteria and fungi was 35.2％ (71 cases),56.4％ (114 cases) and 8.4％ (17 cases),respectively.Comparing with gram negative group and fungi group,gram positive group had older gestational age (36.0 ± 3.8 w,compared with gram negative 33.0 ± 3.9 w,fungi group 31.2 ± 3.2 w,P ＜ 0.05),larger birth weight (2 620 ± 925 g compared with gram negative group 1 999 ± 849 g,fungi group 1 595 ± 666 g,P ＜ 0.05),lower nosocomial infection rate (29.6％ compared with gram negative group 70.2％,fungi group 94.1％,P ＜0.05),lower rate of shock,blood glucose disturbance and thrombocytopenia (P ＜ 0.05).There was no statistics difference between the gram negative group and fungi group.Comparing with the other two groups,fungi group was older [20.0 (11.5,39.5) d compared with gram positive group 7.0 (2.0,17.0) d,and gram negative 10.0 (6.0,18.2) d,P ＜ 0.05].The rate of deep venous catheterization in fungi group was higher than that in gram positive group and gram negative group (88.2％ compared with gram positive group 25.4％,gram negative group 40.4％,P ＜0.05).The treatment course of fungi group was longer than that of the gram positive group and gram negative group [22.0 (12.0,37.5) d compared with the gram positive group 14.0 (10.0,17.0) d,gram negative group 14.0 (11.0,18.0) d,P ＜0.05].The incidence of apnea in the gram negative group was higher than that in gram positive group and fungi group (P ＜ 0.05).The rate of leukocytosis,leukocytopenia and elevated CRP were higher in gram negative and gram positive group (P ＜ 0.05).Conclusion The clinical manifestations and laboratory examinations in neonatal sepsis caused by different pathogens were different,which can help to early identification of different pathogenic infections.However,there is no specific indicators to differentiate neonatal sepsis caused by different pathogens.Early identification of the pathogen needs clinical acumen.

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Non-invasive high frequency ventilation support in the treatment of neonatal respiratory distress syndrome after extubation

Tao ZHANG ; Weiwei GAO ; Jia CHEN ; Xiaorui HUANG ; Yuanhong XIAO ; Shumin KUANG ; Qinghua YANG ; Yunbin CHEN

Chinese Journal of Neonatology.2017;32(2):96-99. doi:10.3760/cma.j.issn.2096-2932.2017.02.005

Objective To compare the efficacy and safety of the non-invasive high frequency ventilation (NHFV) and non-invasive intermittent positive pressure ventilation (NIPPV)in neonates with respiratory distress syndrome (RDS) after extubation.Method Neonates with RDS from January 2015 to January 2016,who required high frequency ventilation after birth and were extubated after treatment were retrospectively studied.The enrolled patients were divided into NHFV group and NIPPV group to compare the rate of successful extubation within 7 days,non-invasive respiratory ventilation support time and complication incidence.Result In total 42 neonates were included,NHFV group were 21 cases and NIPPV group were 21 cases.The rates of successful extubation were not statistically different (71.4％ vs.80.9％,P ＞ 0.05);Compared with NIPPV group,NHFV group had shorter ventilation time [3.5 (2.2,4.1) d vs.4.6 (2.8,5.3)];the incidence of bronchopulmonary dysplasia,pneumothorax,intraventricular hemorrhage,periventricular leukomalacia,retinopathy of premature and necrotizing enterocolitis between groups were not statistically different (P ＞ 0.05).Conclusion NHFV is a new safe and efficient ventilation support method for extubated neonates,and needs further research.

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