Neonatal Necrotizing Enterocolitis Group Research Articles

Efficacy of Probiotic Consortium Transplantation on Experimental Necrotizing Enterocolitis

IntroductionFecal microbiota transplantation (FMT) is a promising therapy, but it has not been used to treat neonatal necrotizing enterocolitis (NEC) due to reports of adverse side effects. Probiotics are considered relatively safe with practicable administrative procedures; however, no systematic research has compared the results of FMT and probiotic consortium transplantation (PCT) on oxidative stress in the intestines of patients with NEC. We conducted this study to provide a basis for optimizing NEC therapy. MethodsEight-day-old newborn C57BL/6 mice were randomly divided into the following four groups: the dam-fed group (control group); the NEC induction group (NEC group); the NEC induction and transplantation of Lactobacillus reuteri and Bifidobacterium infantis consortium group (NEC + PCT group); and the NEC induction and the FMT group (NEC + FMT). Intestinal injury, oxidative stress indexes, intestinal barrier function, and inflammatory cytokines were assessed in the terminal ileum. ResultsFMT more effectively modulates oxidative stress in the intestine than does PCT; however, the difference between the effects of PCT and FMT was not significant. The protective effect was associated with enhanced antioxidant capacity, regulation of the main components of the mucus layer, reduced inflammatory reactions, and improved intestinal integrity. ConclusionsIntestinal dysbiosis affects oxidative stress, inflammatory response, and mucosal integrity. Although FMT is more effective than PCT in regulating oxidative stress, PCT may be preferred in pediatrics because the proportion and dose of transplanted bacteria can be standardized and individualized according to individual conditions.

A Study on Fucosyltransferase 2 Gene Polymorphism and Secretion Status Related to Neonatal Necrotizing Enterocolitis.

Objective To detect the single nucleotide polymorphism (SNP) of alpha-(1,2) fucosyltransferase 2 gene (FUT2) and the secretion status in the newborns of Chongqing China and explore the relationship between genotype or phenotype of FUT2 and neonatal necrotizing enterocolitis (NEC). Methods Newborns who were hospitalized in Children's Hospital of Chongqing Medical University from August 2014 to December 2015 and in line with the inclusion criteria were chosen as the research subjects; 34 cases of them in accordance with the diagnostic criteria of NEC stage II or III were NEC group, and 36 other cases of them were the control group. Total DNA was extracted from oral epithelial cells of patients which were collected with cotton buds. FUT2 SNP genotype was detected by gene sequencing. H antigen was detected with saliva samples by saliva agglutination inhibition test. Related clinical data were collected for analysis. Results There are three genotypes on the rs1047781 (A385T) allele of the FUT2 encoding sequence: AA, AT, and TT. The number of genotypes AA, AT, and TT in the NEC group was 9 (26.47%), 12 (35.29%), and 13 (38.24%), respectively. In the control group, the number of genotypes AA, AT, and TT was 12 (33.33%), 17 (47.23%), and 7 (19.44%). There were no differences in genotypes between the two groups according to the chi-square test (P > 0.05). There were 22 cases of secretors (64.7%) and 12 cases of nonsecretors (35.3%) in the NEC group. The number of secretors and nonsecretors in the control group was 31 (88.89%) and 5 (11.11%). Statistical difference was found in the phenotype between two groups through the chi-square test (P < 0.05). In addition, no G428A homozygous mutation, which causes nonsecretor phenotype in Caucasians, was seen in all the subjects of this study. Conclusions These findings indicate that secretion status (nonsecretor) was significantly associated with NEC in Chongqing, China.

Association of High-Mobility Group Box 1 (HMGB1) Gene Polymorphisms with Susceptibility and Better Survival Prognosis in Chinese Han Neonatal Necrotizing Enterocolitis.

BackgroundHigh-mobility group box 1 (HMGB1) plays a crucial role in a variety of diseases, including neonatal necrotizing enterocolitis (NEC). The purpose of this study was to investigate the association of HMGB1 gene single-nucleotide polymorphisms (SNPs) with susceptibility and survival prognosis in Chinese Han neonates with NEC.Material/MethodsThe HMGB1 gene rs1360485, rs1045411, and rs2249825 site SNPs were genotyped in all participants. The mRNA expression of serum HMGB1 was examined using quantitative reverse transcription-polymerase chain reaction. The correlation of the HMGB1 rs1360485 SNP with NEC neonatal survival prognosis was evaluated by univariate analysis and logistic multivariate regression analysis.ResultsThe TC and CC genotype and C allele distribution frequencies of the rs1360485 SNP were lower in the NEC group, and the differences were statistically significant (all P<0.05). Individuals carrying the TC and CC genotype or C allele had a low risk of being affected by NEC. However, the genotype and allele distributions of rs1045411 and rs2249825 were not significantly different between the patient and control groups (P>0.05). NEC neonates with HMGB1 gene rs1360485 site mutations had lower mRNA levels of serum HMGB1 than those with rs1360485 site wild-type, and the rs1360485 genotypes TC and CC could independently predict better survival outcomes in NEC neonates.ConclusionsThis study demonstrated that the rs1360485 SNP of the HMGB1 gene is associated with susceptibility of NEC in neonates, and the rs1360485 genotypes TC and CC may affect HMGB1 expression and are associated with the survival prognosis of neonates with NEC.

Activated M1 macrophages suppress c-kit expression via TNF-α-mediated upregulation of miR-222 in Neonatal Necrotizing Enterocolitis.

Activation of intestinal macrophages is implicated in the pathogenesis of neonatal necrotizing enterocolitis (NEC), yet its precise mechanisms remain unclear. The purpose of this study is to investigate the role of macrophages and TNF-α via an inflammatory MicroRNA in NEC. Immunofluorescence (IF) staining of CD68, iNOS, and Arg-1 was employed to identify phenotypes of macrophage in the intestines of NEC infants and NEC mice. Expression of TNF-α, c-kit, and miR-222 was evaluated by qRT-PCR, Western blot, and immunochemical staining from the tissue samples. Large number of M1 macrophage infiltration was found in the NEC intestines. Expression of CD68, iNOS, and TNF-α were significantly increased, while c-kit was decreased distinctly in the NEC group. In the early phase of NEC mouse model, inhibition of M1 macrophages reduced the incidence of NEC and intestinal inflammation. We found that TNF-α upregulated the expression of miRNA-222 and inhibited the expression of c-kit. Conversely, such decrease of c-kit expression could be reversed by miR-222 antagonists. Furtherly, dual-luciferase assay confirmed that c-kit can be inhibited by miR-222 directly. Macrophages activation in NEC intestine results in an increased inflammatory response and TNF-α production, accompanied with miR-222 upregulation and c-kit suppression. Modulations of M1 macrophages, TNF-α or miR-222 may be potential therapeutic targets for NEC treatment.

Efficacy of abdominal ultrasound inspection in the diagnosis and prognosis of neonatal necrotizing enterocolitis

OBJECTIVE:This study aimed to identify the most useful ultrasound (US) features associated with definite neonatal necrotizing enterocolitis (NEC) and their prognostic values, particularly the calculated markers combined with important features.METHODS:A total of 213 suspected NEC cases were collected from the neonatal department of our hospital from January 2015 to August 2017. Each infant received both X-ray and US examinations.RESULTS:No differences were found in sex composition and delivery modes between groups. NEC-positive neonates had poorer prognosis compared to negative ones. The NEC group showed a higher frequency of abnormal signals. US showed higher NEC-related frequencies in different parameters. A variable (named predictor in US [PUS]) with five features was constructed. For NEC diagnosis, this variable provided a much higher area under the curve Q2 (AUC) (0.965) than other parameters. In this model, PUS had a cutoff value of 0.376 with a 0.900 sensitivity and 0.922 specificity. In prognosis, the closest factors were selected to draw a receiver operating characteristic curve, as well as a novel calculated variable US prognostic (USPro) marker. USPro had a much higher AUC (0.86) than other single features and showed a cutoff value of 0.18145, with 0.75 sensitivity and 0.84 specificity. This variable had a weaker power in prognosis when compared with PUS in diagnosis.CONCLUSIONS:The application of abdominal color Doppler US can provide high accuracy and sensitivity in NEC diagnosis and also contribute to its prognosis, without induction of radiation. Suspected neonates should be examined using this technique as early as possible.

Changes of intestinal microflora in neonatal necrotizing enterocolitis: a single-center study.

ObjectiveTo investigate the changes in the diversity and dynamics of intestinal microflora in patients with neonatal necrotizing enterocolitis (NEC) in a single center in China.MethodsIn this prospective study conducted between 2016 and 2019, feces were collected from 28 neonates with NEC and 30 control neonates to analyze the species richness and Shannon’s diversity index of the bands, which were also sequenced via PCR-denaturing gradient gel electrophoresis.ResultsThe species richness and Shannon’s diversity index were significantly lower in the NEC group than in the control group, indicating that the diversity of intestinal flora was reduced in NEC. The numbers of samples positive for Bacteroides and Klebsiella were higher in the NEC group, whereas Escherichia coli, Bifidobacterium, and Lactobacillus were less frequently detected in patients with NEC than in controls. The prevalence of intestinal bacteria normalized after the resolution of NEC.ConclusionThe decrease of intestinal flora diversity in neonates with NEC, especially the decreased prevalence of probiotic bacteria, may be an important factor in the pathogenesis of NEC.

Potential role of endoplasmic reticulum stress is involved in the protection of fish oil on neonatal rats with necrotizing enterocolitis

Neonatal necrotizing enterocolitis (NEC) is a serious gastrointestinal disease with high death rate in premature infants. Fish oil (FO) and its constituents have been shown to ameliorate intestinal inflammation and mucosal damage. However, the underlying mechanism of action is not known. In the present study, we divided Sprague-Dawley rats into three groups: control group, NEC model group, and FO pre-feeding+NEC model group. Briefly, one week before NEC modeling, in addition to being fed with milk, the FO pre-feeding+NEC modeling group was fed with FO, the NEC group was fed with saline, and the control group was only inserted a gastric-tube for 7 days. Subsequently, histological assay, Western blot, and ELISA were performed. Pretreatment with FO attenuated the NEC symptoms, alleviated intestinal pathological injury, and decreased the expressions of pro-inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Furthermore, pretreatment with FO reduced the expressions of endoplasmic reticulum stress (ERS) related proteins, caspase-12, and glucose-regulated protein 78 (GRP78). In addition, intestinal histopathological scores showed a significant positive correlation with intestinal expressions of IL-6, TNF-α, and caspase-12. Collectively, these results indicate that ERS pathway might be involved in the effect of FO in alleviating intestinal mucosal inflammation and injury in rats with NEC

Differential diagnosis and treatment of intestinal perforation caused by neonatal necrotizing enterocolitis and Hirschsprung′s disease

Objective To retrospectively analyze the differential diagnosis and treatment strategy of neonatal intestinal perforation caused by neonatal necrotizing enterocolitis (NEC) and Hirschsprung′s disease (HD). Methods The clinical data of 42 cases with NEC and 22 cases with HD combined with intestinal perforation from Ja-nuary 2012 to January 2017 admitted at Fujian Provincial Maternity and Children′s Hospital were collected, the proportion of premature infants, and the proportion of low birth weight infants, age of onset, preoperative clinical symptoms and surgical treatment was compared between the two groups. Results The proportion of premature infants[95.23%(40/42 cases) vs.27.27%(6/22 cases)], the proportion of low birth weight infant[90.48%(38/42 cases) vs.45.45%(10/22 cases)]and the age of onset [(14.48± 10.51) d vs.(3.18±3.43) d]were statistically different between the NEC group and the HD group, and the differences were statistically significant(all P<0.05). There were significant differences in the presence of normal defecation before the operation[71.4% (30/42 cases)vs.27.3%(6/22 cases)], and in the the defecation characteristics at the onset[95.2% (40/42 cases)vs.9.1%(2/22 cases)]between the NEC group and HD group(all P<0.05). Among 42 cases of NEC patients, 8 cases underwent intestinal anastomosis, and 34 cases underwent enterostomy because of the high risk of anastomosis.All of 22 patients with HD underwent double-lumen enterostomy. Conclusions The general conditions, preoperative manifestations and intraoperative fin-dings of the children should be comprehensively analyzed for identification.Enterostomy is recommended for NEC intestinal perforation, but if the risk of anastomosis is high, then fistula should be performed.Double-lumen fistula should be recommended for HD intestinal perforation, and enterostomy should be recommended if the two cannot be clearly distinguished. Key words: Infant, newborn; Intestinal perforation; Necrotizing enterocolitis; Hirschsprung′s disease

Significance of dynamic evolution of TNF-α, IL-6 and intestinal fatty acid-binding protein levels in neonatal necrotizing enterocolitis.

To study the significance of dynamic evolution of serum tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6) and intestinal fatty acid-binding protein (I-FABP) levels in neonatal necrotizing enterocolitis (NEC). A total of 45 NEC child patients, 45 non-NEC child patients and 45 healthy newborns were enrolled. After the day age, weight, gestational week and delivery mode were matched, the serum TNF-α, IL-6 and I-FABP levels at 6, 24 and 72 h after admission were measured via ELISA method, and their correlations with prognosis were analyzed. The levels of serum TNF-α and IL-6 in NEC and non-NEC group reached the peak at 24 h and fell at 72 h; there were no differences in each time point between the two groups (P>0.05), but the levels of serum TNF-α and IL-6 were higher than those in the control group (P<0.05). The level of serum I-FABP in NEC and non-NEC group reached the peak at 6 h, and it fell at 72 h in NEC group and 24 h in non-NEC group; the level of I-FABP in each time point in NEC was significantly higher than that in non-NEC group, and the level was the lowest in healthy group; the differences were statistically significant (P<0.05). There were 40 cases of survival and 5 cases of death (11.1%) in NEC group, while there were 43 cases of survival and 2 cases of death (4.4%) in non-NEC group. There were no differences in serum TNF-α and IL-6 levels at different times between surviving child patients and dead child patients in NEC group (P>0.05), but the levels of serum I-FABP in surviving child patients at 6 h and 24 h were significantly lower than those in dead child patients (P<0.05), and there was no difference at 72 h (P>0.05). There were no differences in serum TNF-α, IL-6 and I-FABP levels at different times between surviving and dead child patients in non-NEC group (P>0.05). Serum I-FABP level and its dynamic evolution may be important indexes of early diagnosis and prognosis evaluation of NEC.

Differential expression of microRNA in neonatal necrotizing enterocolitis based on high-throughput sequencing

Objective To analyze the differential expression of microRNA (miRNA) and its significance in patients with neonatal necrotizing enterocolitis (NEC). Methods Twenty-five patients diagnosed with NEC with Bell stage ≥Ⅱ, and 25 non-NEC patients as control group admitted to Guangzhou Women and Children's Medical Center between October 2014 and November 2015 were collected. White blood cells were extracted from the peripheral blood. Five samples were selected randomly each from NEC group and control group, and sequenced by second-generation Illumina high-throughput sequencing, screened for differentially expressed miRNA and analyzed for target genes prediction and biological function. The rest samples of the two groups were detected by real-time fluorescent quantitative polymerase chain reaction technology (RT-qPCR), the results were used to validate the results of high-throughput sequencing. Differentially expressed miRNA in the two groups of data was analyzed using DEGseq software. P<0.05 was considered statistically significant. P<0.01, q<0.001 and丨Log2 Ratio丨≥1 were taken as criteria for screening the differential expression. The differential expressions of miRNA in NEC group and control group were analyzed by cluster analysis using MeV4.6 software. Results A total of 482 miRNAs were differentially expressed in the two groups, with significant difference (P<0.05). Among them, 126 were known miRNAs with significantly differential expression in the two groups, with 58 being up-regulated and 68 being down-regulated. The results of up-regulated miRNAs (hsa-miR-223-5p, -183-3p, -222-5p) and down-regulated miRNAs (hsa-miR-23b-5p, -150-5p, -146a-3p, -1298-5p) were confirmed to be consistent with the results of sequencing. Bioinformatics analysis showed that the target genes with differential miRNA expression mainly involved Toll-like receptor signal transduction pathway, mitogen-activated protein kinase pathway, JAK-STAT and other signal transduction pathways. Conclusions There are significantly differential expressions of miRNAs in peripheral white blood cells of NEC neonates. These miRNs may be involved in the occurrence and development of NEC via adjusting different target genes to regulate the signal pathway. Key words: Enterocolitis, necrotizing; MicroRNAs; High-throughput nucleotide sequencing; Infant, newborn

Value of abdominal ultrasound in the diagnosis of neonatal necrotizing enterocolitis and evaluation of disease severity

To investigate the value of abdominal ultrasound in diagnosing neonatal necrotizing enterocolitis (NEC) and its significance in evaluating the disease severity. The clinical data of 84 neonates who were diagnosed with NEC between July 2013 and January 2015 were analyzed retrospectively. According to the modified Bell-NEC staging criteria, these neonates were divided into a suspected NEC group (n=44) and a confirmed NEC group (n=40); according to clinical prognosis, they were divided into a medical treatment and full recovery group (n=58) and a surgery/death group (n=26). The changes in the results of abdominal ultrasound and abdominal X-ray plain film were compared between groups. In the confirmed NEC group, abdominal ultrasound showed significantly higher detection rates of portal venous gas and dilatation of the intestine than abdominal X-ray plain film (P<0.05). Compared with the medical treatment and full recovery group, the surgery/death group had significantly higher detection rates of dilatation of intestine, bowel wall thickening, peritoneal effusion and free intraperitoneal air (P<0.05). Dilatation of the intestine and free intraperitoneal air shown by abdominal X-ray plain film were more common in the surgery/death group. Abdominal ultrasound is useful for the diagnosis of NEC. Ultrasonic findings can contribute to the prediction of the severity of NEC.

Diagnostic value and disease evaluation significance of abdominal ultrasound inspection for neonatal necrotizing enterocolitis.

Objective:To summarize abdominal plain X-rays and ultrasound characteristics of 144 cases of Neonatal Necrotizing Enterocolitis (NEC) and to analyze diagnostic value and disease evaluation significance of abdominal ultrasound inspection for NEC.Methods:Clinical data of 144 NEC patients were retrospectively analyzed from February 2014 to December 2015. The patients were divided into suspected NEC group (N=74) and confirmed NEC group (N=70) according to amended Bell-NEC classification and diagnostic criteria. Meanwhile, we divided them into internal medicine treatment group (N=95) and surgery/death group (N=49) according to clinical prognosis and took records of their clinical manifestations, laboratory inspection results and abdominal plain X-rays and ultrasound characteristics.Results:For confirmed NEC group, the detection rate of portal venous gas (PVG) and dilatation of intestine by abdominal ultrasound was obviously higher than by plain X-rays (P<0.05). Abdominal ultrasound inspection revealed that the incidence rate of dilatation of intestine, bowel wall thickening and ascites (acoustic transmission difference) of the surgery/death group was higher than that of the internal medicine treatment group by comparing risk ratio (RR) and 95% confidence interval (CI) of RR; the difference was statistically significant (P<0.05). The abdominal plain X-rays inspection only showed the result that dilatation of intestine and free intraperitoneal air was more often found in the surgery/death group (P<0.05).Conclusion:Compared with abdominal plain X-rays, abdominal ultrasound has certain clinical value and offers more advantages in some aspects; therefore, it can be considered as the reference index in prediction of clinical prognosis.

Association of neonatal necrotizing enterocolitis with myeloid differentiation-2 and GM2 activator protein genetic polymorphisms.

The aim of the present study was to investigate the association of neonatal necrotizing enterocolitis (NEC) with myeloid differentiation-(MD-2) and GM2 activator protein (GM2A) genetic polymorphisms. Gene resequencing of the MD-2 and GM2A gene exons was performed on 42 neonates, diagnosed with NEC (NEC group), as well as in the rs 11465996 locus, located in the MD-2 gene promoter region. The aim was to detect the genetic polymorphisms present in the neonates with NEC and compare the functional polymorphic loci with 83 neonates without NEC (control group), who had been born during the same period. A polymorphic locus with abnormal frequency was detected in the exon region of the MD-2 gene. In the NEC group, the frequency of genotypes carrying the low frequency allele (G) in the rs 11465996 locus (MD-2 promoter region) was significantly higher compared with the control group (χ2=4.388, P=0.036). Furthermore, the frequencies of genotypes carrying the low frequency A and C alleles in the rs1048719 (GM2A gene exon 1) and rs2075783 loci (GM2A intron), respectively, were significantly higher in the NEC group compared with the control group (χ2=4.316, P=0.038; and χ2=13.717, P=0.000, respectively). In addition, the rs 11465996 polymorphism in the MD-2 gene promoter region was found to be associated with the severity of NEC. Furthermore, the rs2075783 polymorphism in the GM2A gene exon 1 and the rs1048719 polymorphism in the intron region of this gene, were associated with the occurrence of NEC. The present study demonstrated that gene polymorphisms of MD-2 and GM2A were associated with the occurrence or severity of NEC; however, further in-depth exploration is required to clarify the associations between genetic predispositions to polymorphisms, and NEC.

Effects of heparin-binding epidermal growth factor-like growth factor on mitochondrial pathway of apoptosis in neonatal rats with necrotizing enterocolitis

Objective To investigate the effect of heparin-binding epidermal growth factor-like growth factor (HB-EGF) on mitochondrial pathway of apoptosis in rats with neonatal necrotizing enterocolitis (NEC).Methods Sprague-Dawley neonatal rats were randomly divided into three groups with ten in each.NEC group rats were formula fed,and hypoxia exposed by 100％ N2 for 90 s and cold stress at 4 ℃ for 10 min twice a day for three days.Additionally,rats in HB-EGF group received HB-EGF 800μg/kg by gavage four times a day for three days.Rats in control group were given breast milk feeding for three days without any interventions.Seventy-two hours after born,all neonatal rats were sacrificed after fasting for 12 h,from which the terminal ileum was removed.HE-staining was done for histologic evaluation.Mitochondrial ultrastructure was observed under electron microscopy.Cytochrome C was detected by immunohistochemical analysis and apoptosis inducing factor (AIF) and apoptotic protease activating factor-1 (APAF-1) were measured by Western blot.Analysis of variance and q-test were used to compare the difference among groups.Results (1) The incidence of NEC in HB-EGF group was lower than that in NEC group (2/10 vs 9/10,x2 =7.27,P＜0.01).(2) In NEC group,mitochondria in epithelial cells and muscle cells of intestine were significantly swelling,appearing many electron-lucent zones in matrix.Ultrastructure of mitochondria were severely damaged.In HB-EGF group,mitochondria were less swelling and showed milder damage than those in NEC group.(3) The expression of cytochrome C in ileal tissue in NEC group was higher than that in control group (0.030±0.018 vs 0.002±0.001,q=6.15,P＜0.01).The expression of cytoehrome C in ileal tissue in HB-EGF group was lower than that in NEC group (0.014±0.018 vs 0.030±0.018,q=3.53,P＜0.05).The expression of APAF-1 and AIF in NEC group was higher than those in control group (1.364±0.299 vs 0.215±0.033,q=15.31,P＜0.05;0.181±0.050 vs0.127±0.045,q=3.71,P＜0.05).Compared to NEC group,the expression of APAF-1 was lower (0.455±0.123 vs 1.364±0.299,q=4.04,P＜0.05) and the expression of AIF was higher (0.289±0.045 vs 0.181±0.050,q=7.32,P＜0.05) in HB-EGF group.Conclusions HB-EGF could reduce the incidence of NEC in neonatal rats by inhibiting the mitochondrial pathway related apoptosis through down regulation of APAF-1. Key words: Enterocolitis, necrotizing; Intercellular signaling peptides and proteins; Apoptotic protease-activating factor 1 ; Apoptosis ; Mitochondria ; Rats

Relationship between plasma D-lactate levels and prognosis of neonatal necrotizing enterocolitis

Objective To investigate the relationship between plasma D-lactate levels and prognosis in newborns with neonatal necrotizing enterocolitis (NEC).Methods One hundred and four premature infants with NEC (stase Ⅱ and Ⅲ ) and another 104 premature infants without NEC admitted into Quanzhou Children's Hospital for other diseases from January 2007 to October 2010 were selected into this case control study.The gestational age,gender and birth weight of patients in the two groups were matched.NEC patients' bloods were collected within 24 hours after NEC was confirmed and blood samples of the control group were collected at the corresponding age.Enzymelinked immunosorbent assay was used to detect the plasma D-lactate level.Receiver operating characteristic curve was used to confirm the criteria of D-lactate positive,according to which,the NEC group was divided into high D-lactate group and normal D-lactate group,and the mortality and complication rate of the two groups were compared with x2 test.And NEC group was subdivided into death group and survive group according to the prognosis of the patients,and the difference of D-lactate level between the two groups were compared with t test.Results Among the NEC group,there were 63 cases (60.6％) of stage Ⅱ and 41 (39.4％) cases of stage Ⅲ ; 88 (84.6％) survived infants and 16 (15.4％) dead infants.D-lactate level was (35.4 ± 29.1) μg /ml in stage Ⅲ NEC group,(29.5±16.2) μg/ml in stage Ⅱ NEC group and (3.7±18.4) μg/ml in control group; there were statistical differences between each other(F=5.97,P＜0.05).Receiver operating characteristic curve analysis showed that if 6 μg/ml was set as the borderline,there were 87 cases(83.7％,87/104) with high D-lactate patients in NEC group,whose neonatal critical illness scoring (NCIS) was 80.9±22.6,significantly lower than that of normal D-lactate patients ( 95.8 ± 20.5) (t =2.417,P＜ 0.05),and higher neonatal septicemia rate (48.3％,42/87) and mortality (27.6％,24/87) compared with those [(5.9％,1/17) and (5.9％,1/17)] of normal D-lactate patients(x2 =11.539 and 7.146,P＜0.05,respectively).In NEC group,compared with the survived infants,the D-lactate level [(43.2±13.5) μg/ml vs (21.9 ± 22.9) μg/ml,t =4.572,P＜0.05] and the rate of septicemia [68.8％ (11/16) vs 38.6％ (34/88),x2 =3.445,P＜0.05] were higher in dead patients,and NCIS (82.4± 29.1 vs 90.6 ± 21.3,t =2.409,P＜0.05) was lower.Conctusions The level of plasma D-lactate related to the prognosis of neonatal NEC and which might be a good indicator for its prognosis and severity. Key words: Enterocolitis, necrotizing; Lactic acid; Infant, newborn; Prognosis

Captopril reduces the severity of bowel damage in a neonatal rat model of necrotizing enterocolitis

Selective mesenteric ischemia may result from activation of the renin-angiotensin system during periods of shock and is implicated in the pathogenesis of neonatal necrotizing enterocolitis (NEC). We investigated the effectiveness of captopril, an angiotensin-converting enzyme inhibitor, in reducing the severity of bowel damage in a neonatal rat model of NEC. Necrotizing enterocolitis was induced by a combination of gavage feeding of hypertonic formula, hypoxia, and oral lipopolysaccharide (LPS). Rats were randomly divided into 3 groups: group A, control (breast fed; n = 20); group B, NEC (gavage/hypoxia/LPS; n = 31); group C, NEC with captopril 20 mg/kg per dose with the formula for 4 days (gavage/hypoxia/LPS/captopril; n = 35). Pups were killed after 4 days. Incidence of NEC was evaluated microscopically. Severity of bowel damage was higher in the NEC group compared to controls and was reduced by administration of captopril. Dilatation of the intestinal vasculature was observed in the captopril group. There were no cases of NEC in the controls; the incidence increased to 55% in NEC group and reduced to 29% by captopril. In this model of neonatal NEC, captopril supplementation of formula reduces the severity of intestinal damage and the incidence of NEC, presumably by affecting mesenteric blood flow.

Epidermal Growth Factor Reduces Hepatic Sequelae in Experimental Necrotizing Enterocolitis

Background and Aim: Neonatal necrotizing enterocolitis (NEC) is the most common gastrointestinal disease of premature infants. We recently demonstrated that the gut/liver axis plays an important role in the pathophysiology of NEC through the release of inflammatory mediators into the intestinal lumen. We have also shown that supplementation of formula with epidermal growth factor (EGF) dramatically decreases ileal pathology associated with experimental NEC. In this study, we examined the effects of EGF on the liver portion of the gut/liver axis in the neonatal rat model of NEC. Methods: Newborn rats were divided into three experimental groups, NEC, hand-fed with growth-factor free formula; NEC + EGF, hand-fed with formula supplemented with 500 ng/ml rat EGF; or DF, dam fed. All animals were exposed to asphyxia and cold stress twice daily for 4 days to develop NEC. Results: EGF receptor expression was significantly (p ≤ 0.01) decreased in the NEC + EGF group compared to the NEC group. EGF supplementation significantly decreased Kupffer cell numbers (p ≤ 0.01) as well as hepatic tumor necrosis factor (TNF)-α and interleukin-18 production (p ≤ 0.05). Further, TNF-α in the intestinal luminal contents of the NEC + EGF group were normalized to levels observed in DF controls compared to the NEC group (p ≤ 0.05). Activated nuclear factor-ĸB was also substantially decreased in the NEC + EGF group versus the NEC group. Conclusion: The results of this study indicate that EGF normalizes cytokine overproduction in the liver of neonatal rats with NEC, which contributes to diminished intestinal damage during the development of experimental NEC. These data suggest that supplementation of formula with EGF can have beneficial effects on the gut/liver axis during NEC pathogenesis.

Diminished epidermal growth factor levels in infants with necrotizing enterocolitis

Background/Purpose: Because epidermal growth factor (EGF) is trophic to the intestinal mucosa, and neonatal necrotizing enterocolitis (NEC) is associated with a disrupted intestinal mucosal barrier, the authors sought to determine whether diminished levels of EGF were present in infants with NEC.Methods: Saliva, serum, and urine specimens were obtained from infants with NEC during a 3-year period (February 1995 to May 1998). Control patients without NEC were chosen based on similar postnatal age and birthweight. EGF levels were determined by enzyme-linked immunosorbent assay (ELISA). Differences between groups were compared using Mann-Whitney Rank sum test with P less than .05 considered significant. Results are presented as mean values ± SEM.Results: Twenty-five infants with NEC were compared with 19 control patients. Birth weight (1,616 ± 238 g control v 1,271 ± 124 g NEC) and postnatal age (23 ± 6 days control v 22 ± 3 days NEC) were similar. Infants with NEC had significantly lower levels of EGF in both saliva (590 ± 80 pg/mL control v 239 ± 41 pg/mL NEC; P < .001) and serum (35 ± 8 pg/mL control v5.6 ± 1.9 pg/mL NEC; P < .001). Urinary EGF was also lower in the NEC group, but was not statistically significant.Conclusions: Premature infants with NEC have significantly diminished levels of salivary and serum EGF. Reduced levels of this growth factor may distinguish infants at risk for NEC and play a pivotal role in the pathogenesis of the perturbed intestinal mucosal barrier that is central to this condition.

ASSOCIATION OF MESENTERIC THROMBOEMBOLISM AND ORAL FEEDINGS WITH NEONATAL NECROTIZING ENTEROCOLITIS

Splanchnic ischemia is believed to be a major factor in the development of neonatal necrotizing enterocolitis (NEC). The role of vascular occlusion in the pathogenesis of NEC was studied in 30 autopsies. Multiple histologic sections of the mesentery were examined for the presence of thrombotic phenomena. Evidence of systemic thrombosis was also sought. 16 infants had a postmortem diagnosis of NEC (Group A) and 14 infants had intestinal hemorrhage or congestion (Group B). The clinical course of each patient was reviewed. 15/16 in Group A received oral feedings and 5/14 in Group B (P < .001). In Group A 12/15 had thromboemboli in mesenteric muscular arteries (TMA). 10/16 had evidence of multiple thromboemboli in other organs: kidney 7, adrenal 4, lung 4, and liver 2. 11/13 had mural thrombi of the abdominal aorta and iliac arteries. 15 had umbilical artery catheters (UAC) and 8 had umbilical venous catheters (UVC). In Group B 2/13 had TMA (P< .001). 11/14 had multiple thromboemboli in other organs: kidney 4, adrenal 3, lung 7, liver 2, and spleen 5. 9/12 had mural thrombi of the abdominal aorta and iliac arteries. 11 had UAC and 7 had UVC. There is an apparent association of NEC with meaenteric thromboembolism and oral feedings in this study.

ASSOCIATION OF MESENTERIC THROMBOEMBOLISM AND ORAL FEEDINGS WITH NEONATAL NECROTIZING ENTEROCOLITIS

Splanchnic ischemia is believed to be a major factor in the development of neonatal necrotizing enterocolitis (NEC). The role of vascular occlusion in the pathogenesis of NEC was studied in 30 autopsies. Multiple histologic sections of the mesentery were examined for the presence of thrombotic phenomena. Evidence of systemic thrombosis was also sought. 16 infants had a postmortem diagnosis of NEC (Group A) and 14 infants had intestinal hemorrhage or congestion (Group B). The clinical course of each patient was reviewed. 15/16 in Group A received oral feedings and 5/14 in Group B (P < .001). In Group A 12/15 had thromboemboli in mesenteric muscular arteries (TMA). 10/16 had evidence of multiple thromboemboli in other organs: kidney 7, adrenal 4, lung 4, and liver 2. 11/13 had mural thrombi of the abdominal aorta and iliac arteries. 15 had umbilical artery catheters (UAC) and 8 had umbilical venous catheters (UVC). In Group B 2/13 had TMA (P< .001). 11/14 had multiple thromboemboli in other organs: kidney 4, adrenal 3, lung 7, liver 2, and spleen 5. 9/12 had mural thrombi of the abdominal aorta and iliac arteries. 11 had UAC and 7 had UVC. There is an apparent association of NEC with meaenteric thromboembolism and oral feedings in this study.