Insulin resistance biomarkers in small-for-gestational-age infants born to mothers with gestational diabetes mellitus.

OBJECTIVE: Early alterations in glucose homeostasis increase the risk of developing insulin resistance (IR) and obesity later in life. The study aimed to ascertain the peripheral blood levels of hormones that controlling glucose homeostasis and inflammatory factors that are correlated with IR and fetal outcomes in small-for-gestational-age (SGA) infants born to mothers with gestational diabetes mellitus (GDM). METHODS: This cohort study included a total of 90 SGA infants born to mothers with GDM (n = 37) and without GDM (n = 53). At birth, blood levels of glucose, insulin, C-peptide, growth hormone (GH), IGFBP3, lipid profiles, fibrinogen, and hypersensitive C-reactive protein (Hs-CRP) were measured; homeostatic model assessment-IR (HOMA-IR) and ponderal index were calculated. All newborns were followed up to the first year of life. RESULTS: Compared with SGA infants born to mothers without GDM, the levels of low-density lipoprotein-cholesterol (LDL-C), GH, and fibrinogen were significantly higher in the SGA infants born to mothers with GDM (p = .048, .045, and .04, respectively). However, total cholesterol, HDL-C, and apolipoprotein(a) levels were significantly lower in the SGA infants born to mothers with GDM when compared with those in with SGA infants born to mothers without GDM (all p < .05). Weight gain in the first year was higher in the SGA infants born to mothers with GDM group than SGA infants born to mothers without GDM [6644 g (5991-7572) vs. 6032 g (5529-6932)]. CONCLUSIONS: Altered biomarkers of IR were observed among SGA infants born to mothers with GDM, suggesting that these infants were more prone to develop IR after birth.

Demographic and Clinical Features of Small-for-Gestational-Age Infants Born to Mothers With Gestational Diabetes Mellitus.

We studied the demographic and clinical characteristic, risk factors, outcomes of full-term small-for-gestational-age (SGA) infants born to mothers with gestational diabetes mellitus (GDM) in China. A retrospective case-control study that included 1981 SGA infants was conducted; the demographic and clinical data between SGA infants born to mothers with and without GDM were compared. Of 383 SGA infants born to mothers with GDM, 221 (57.7%) were female, and the incidence of these infants was 1 in 155 live births. The risk of SGA siblings (RR, 1.88; 95% CI, [1.23-2.86]), low 1- and 5-min Apgar scores (RR,2.04 and 4.21; 95%CI [1.05-4.00] and [1.05-16.89], respectively), early thrombocytopenia (RR, 3.39; 95%CI, [1.33-8.64]), hypoglycemia(RR, 2.49; 95%CI, [1.55-3.98]), and hypoxic-ischemic encephalopathy (RR,5.61; 95%CI, [1.25-25.18]) were increased in SGA infants born to mothers with GDM compared to SGA infants born to mothers without GDM. SGA girls born to mothers with GDM had a significantly higher ratio of catch-up growth (CUG) (RR, 1.73; 95%CI, [1.18-2.54]) in the first year of life. These results show that genetic factors may be one of the etiologies of SGA infants born to mothers with GDM; and these infants have more adverse perinatal outcomes compared to SGA infants born to mothers without GDM. SGA girls born to mothers with GDM had accelerated CUG in the first year of life.

Prader-Willi syndrome in neonates: twenty cases and review of the literature in Southern China.

BACKGROUND: Prader-Willi syndrome is a rare genetic abnormality that can be challenging to diagnose early, but for which early interventions improve prognosis. METHODS: To improve understanding of Prader-Willi syndrome in neonates in Asia, we retrospectively analyzed the clinical records of 20 affected newborns diagnosed in the Department of Neonatology, Guangzhou Women and Children's Medical Center, Guangzhou, China from January 2007 to December 2014 and performed a review of the relevant literature. RESULTS: Fourteen boys and six girls presented with hypotonia, poor responsiveness, feeding difficulty, and infrequent, weak crying. Different from western patients, the 20 Asian patients exhibited at least five of the following typical features: prominent forehead, narrow face, almond-shaped eyes, small mouth, downturned mouth, thin upper lip, and micromandible. All 14 boys had a small scrotum, including nine with cryptorchidism. Diagnoses were made with microarray comparative genomic hybridization. All 20 infants required feeding tubes. Fifteen received swallowing training immediately after admission; the period of continuous tube feeding for these patients ranged from 8 to 22 days (mean, 14 ± 5.3 days). For the five patients who did not receive swallowing training, the period of continuous tube feeding ranged from 15 to 35 days (mean, 18 ± 4.3 days). Comprehensive care measures included: giving parents detailed health education and basic information about this disease, teaching skills to promote feeding and prevent suffocation, increasing children's passive activity, providing nutrition management for normal development, and preventing excessive or inadequate nutrient intake. CONCLUSIONS: Neonates with Prader-Willi syndrome in Asia have hypotonia, poor responsiveness, feeding difficulty, infrequent and weak crying, genital hypoplasia, and characteristic facial features. Recognition of the syndrome in neonates with confirmation by genetic testing is essential, because early diagnosis allows early intervention. Treatment measures including swallowing training can improve prognosis, prevent growth retardation and obesity, and elevate quality of life in individuals with Prader-Willi syndrome.

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 rs11465996 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 rs11465996 locus (MD-2 promoter region) was significantly higher compared with the control group (χ(2)=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 (χ(2)=4.316, P=0.038; and χ(2)=13.717, P=0.000, respectively). In addition, the rs11465996 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.

Glutamine downregulates TLR-2 and TLR-4 expression and protects intestinal tract in preterm neonatal rats with necrotizing enterocolitis.

BACKGROUND/PURPOSE: Toll-like receptor (TLR)-4 and TLR-2 play an essential role in the pathogenesis of necrotizing enterocolitis (NEC). In this study, we investigated the protective effect of glutamine (Gln) in an NEC neonatal rat model, and the potential association with TLR-4 and TLR-2 expression in local intestinal tissues. METHODS: Preterm neonatal rats were randomly divided into 3 groups: normal control; NEC model; and NEC plus Gln intervention. NEC was induced by feeding with artificial milk substitutes, plus exposure to hypoxia and cold stress. All preterm rats were sacrificed at 3 days after birth. The intestinal tissues were taken for pathological analysis. Protein and mRNA expression of TLR-2, TLR-4, and caspase-3 was examined by immunohistochemistry and real-time RT-PCR, respectively. RESULTS: Compared with the normal control, the NEC neonatal rats showed mucosal injury and upregulated mRNA and protein expression of TLR-2, TLR-4, and caspase-3 in ileum and colon. Gln intervention significantly reduced the mucosal injury and suppressed the upregulated expression of TLR-2, TLR-4, and caspace-3 in the ileum and colon of NEC neonatal rats. CONCLUSIONS: Gln protects the intestinal tract of NEC neonatal rats, which may be associated with the reduction of TLR-2 and TLR-4 expression in intestines.

[Effects of glycomacropeptide on damage to intestinal tissue and apoptosis of intestinal epithelial cells in neonatal rats with necrotizing enterocolitis].

OBJECTIVE: To establish an appropriate neonatal rat model of necrotizing enterocolitis (NEC) and to investigate the protective effects of glycomacropeptide (GMP) on the gut from injury in neonatal rats with NEC. METHOD: A total of 36 neonatal SD rats were randomly divided into 3 groups: NEC model group (Group M), NEC + GMP group (Group G) and normal control group (Group N), each group had 12 rats. All the neonatal rats were fed with breast milk in the first 3 days after birth. During the second 3 days after birth, the rats of Group N were still maternal breast-fed, but the rats of Group M and Group G were separated from their mothers and lived in incubator and began to be formula fed, and were subjected to cold exposure shortly after hypoxic-reoxygenation treatment. After being fed in such means for 6 days, all the neonatal rats were placed into the incubator and fasted for 24 hours. Then all the rats were sacrificed by cervical dislocation. Intestinal tissue located at the boundary of ileum and cecum was obtained for: (1) histological examination after HE staining, (2) TUNEL detection, (3) electron microscopic observation; and the tissue homogenate was obtained for checking TNF-α and IL-1ß levels by ELISA and platelet activating factor (PAF) mRNA expression by quantitative fluorescence (QF)-PCR. RESULT: (1) The pathological scores of the 3 groups were 2.17 ± 0.83 (Group M), 0.92 ± 0.79 (Group G) and 0.17 ± 0.39 (Group N) separately. There was significant difference between Group M and Group G (H = 8.819, P = 0.003). (2) TNF-α levels of 3 groups were (41.94 ± 13.51) pg/ml (Group M), (31.69 ± 11.68) pg/ml (Group G) and (17.42 ± 7.18) pg/ml (Group N) separately, and TNF-α level in Group G was significantly lower than that of Group M (F = 3.959, P = 0.030). (3) IL-1ß levels of 3 groups were (150.33 ± 36.41) pg/ml (Group M), (118.36 ± 33.00) pg/ml (Group G) and (28.44 ± 15.04) pg/ml (Group N) separately, and IL-1ß level in Group G was lower than that of Group M (F = 5.080, P = 0.013). (4) Expression levels of intestinal PAF mRNA (2(-ΔΔCt) value): 3.01 ± 0.96 (Group M), 1.56 ± 0.29 (Group G), 1.01 ± 0.13 (Group N), the level of Group G was significantly lower than that of Group M (F = 25.251, P = 0.000). (5)Electron microscopy: Group N showed that its cell volume was mostly occupied by the nucleus, the structure was clear, nuclear membrane existed, suggesting the normal phase of cell; Group M showed that apoptotic body existed, suggesting that the advanced stage phase of apoptosis; Group G showed that condensed chromatin marginated around the nuclear envelope, nuclear pores expanded, suggesting the early phase of apoptosis. (6) The apoptosis rate of intestinal epithelial cells by TUNEL detection: 38.79 ± 9.79 (Group M), 29.54 ± 7.30 (Group G), 6.37 ± 1.96 (Group N); the apoptosis rate of intestinal epithelial cells of Group G was significantly lower than that of Group M (F = 6.888, P = 0.003). CONCLUSION: GMP has protective effects on guts of neonatal rats with NEC, which may probably work by reducing TNF-α, IL-1ß and PAF expression, inhibiting the apoptosis of intestinal epithelial cells and reducing intestinal tissue injury.

Establishment and evaluation of three necrotizing enterocolitis models in premature rats.

Necrotizing enterocolitis (NEC) is a gastrointestinal disease that usually affects premature infants and has high morbidity and mortality rates. Reliable animal models aid further study of the etiological factors, pathogenesis, prevention and treatment of NEC. The present study aimed to establish NEC models in premature rats using three common methods, and to determine the optimal model establishment method. The study consisted of six groups; in group A, rats were raised with simulated milk and subjected to inhalation of 100% nitrogen gas (hypoxia) for 90 sec, followed by exposure to cold ambient conditions (4˚C) for 10 min twice daily for 3 days. In group B, rats were exposed to 100% nitrogen gas for 5 min and 100% oxygen for 5 min twice daily for 3 days. Group C rats were intraperitoneally administered 5 mg/kg lipopolysaccharide. Group D and E rats did not receive any intervention. Group F rats were intraperitoneally administered 1 ml/kg physiological saline. Groups D-F served as the control groups corresponding to groups A-C, respectively. Following hematoxylin and eosin staining, intestinal tract, liver, lung and kidney tissues were observed under optical microscopy and were scored. Successful NEC induction was measured by a score of ≥2. Rats from groups A-C exhibited reduced movement, abdominal distention, diarrhea, intestinal tract expansion, and congestion to varying degrees. The pathological scores of intestinal injury in groups A-F were 3.13±0.64, 1.40±0.52, 2.00±0.42, 0.30±0.48, 0.30±0.48, and 0.40±0.52 points, respectively. Significant differences were found between the model groups and their corresponding control groups (p<0.01). Among the model groups, the histological score of group A was higher than that of groups B (p<0.01) and C (p<0.05). The morbidity rate of NEC in groups A-C was 75, 20 and 50%, respectively. There was no morbidity in groups D-F. Compared with groups A and B, injury to the liver, kidney and lung was more severe in group C. Similar symptoms were not observed in groups D-F. Compared with methods of simple hypoxia-reoxygenation or intraperitoneal administration of lipopolysaccharide, the combination of artificial feeding and hypoxia plus cold stimulation most resembles the pathological causes of neonatal NEC. This method resulted in high morbidity, reproducibility and specificity, and was therefore considered an ideal model for establishing NEC.

[Regulatory effects of glutamine on Toll-like receptor 4 in neonatal rats with necrotizing enterocolitis].

OBJECTIVE: To study the expression of Toll-like receptor 4 (TLR-4) and caspase-3 in the intestine of neonatal rats with necrotizing enterocolitis (NEC), and explore the protective effects and possible regulatory mechanisms of glutamine (Gln) in NEC. METHODS: Sixty premature rats were randomly divided into three groups (n=20 each): control, NEC model and Gln intervention group. NEC model was prepared by formula feeding, hypoxia and cold stress. The Gln intervention group was also subjected to hypoxia and cold stress but was fed with formula containing Gln (0.3 g/kg). Two days later, the rats were sacrificed and the intestine tissues were obtained. The histological changes of ileal tissues were observed by hemetoxylin and eosin staining. The expression of caspase-3 and TLR-4 protein in the jejunum, ileum and colon were detected by inmunohistochemistry. The expression of TLR-4 mRNA in the jejunum, ileum and colon were detected by RT-PCR. RESULTS: Compared with the control group, the histological score of ileal tissues, and the expression of caspase-3, TLR-4 protein and TLR-4 mRNA in the NEC model group increased significantly (P<0.01). Gln intervention decreased significantly the histological score of ileal tissues, and the expression of caspase-3, TLR-4 protein and TLR-4 mRNA compared with the NEC model group (P<0.05). CONCLUSIONS: TLR-4 might be involved in the pathogenesis of NEC. Gln may provide protective effects on intestine possibly through reducing the TLR-4 expression and then decreasing the apoptosis of intestinal epithelial cells.