Analysis of serum metabolism in premature infants before and after feeding using GC-MS and the relationship with necrotizing enterocolitis.

Preterm birth and enteral feeding are two main factors leading to necrotizing enterocolitis (NEC). The metabolomics of preterm infants before and after feeding can provide a basis for the prediction of NEC. Using the method of cross-sectional study, the mode was established with the serum samples of 19 premature infants at birth and after feeding as the control group. The serum was analyzed using GC-MS. Chemometric analysis includes principal component analysis, partial least squares-discriminant analysis, and orthogonal partial least squares-discriminant analysis. Spectral separation of serum metabolites occurred in premature infants before and after feeding. The levels of xylose, d-talose, phosphoglycolic acid, maleimide, l-gulonolactone, maleic acid, ß-hydroxypyruvate, itaconic acid, and pantothenic acid in the serum of premature infants after feeding were significant in both multidimensional and single-dimensional modes (variable importance in projection >2, P < 0.01). There was a moderate correlation between total bilirubin and l-gulonolactone and ß-hydroxypyruvate (0.8 > r > 0.5). Maleimide, maleic acid, and itaconic acid have diagnostic value (area under the curve >0.9). The results indicated that serum metabolism of preterm infants changes significantly after feeding. Some metabolites have potential value in predicting NEC.

Intestinal tract and parenteral multi-organ sequential pathological injury caused by necrotizing enterocolitis.

BACKGROUND: To explore the relationship between the pathological changes of the colon, terminal ileum, lung, liver and kidney, and the changes of Bax, PCNA and PAF in a rat model of NEC. METHODS: One hundred and forty neonatal SD rats were randomly divided into NEC group and control group (70 in each group). NEC group was given hypoxia, cold stimulation and artificial feeding twice a day for 3 consecutive days. The control group was only fed normally. After modeling, From the 1st day to the 7th day, 10 rats were sampled in each group for pathological examination of colon, terminal ileum, lung, liver and kidney tissue. The levels of Bax, PCNA and PAF were investigated by immunohistochemistry. RESULTS: Compared with the normal group, in the NEC group, on the 1st day, the colon, terminal ileum, lung, liver and kidney showed inflammatory damage. On the 5th day, the inflammatory injury was reduced. The inflammation disappeared on the 7th day. There were differences in the time of apoptosis in the intestine. In the intestine, the proliferation of PCNA was weak at first and then strong. Bax in liver and kidney showed marked apoptosis and apoptosis time increased in the lung. The expression of PCNA increased in lung, liver and kidney, and the expression of PAF increased in lung and liver. CONCLUSIONS: NEC can lead to secondary injury of different degrees in colon, terminal ileum, lung, liver and kidney, and the degree and time of injury and repair were different. In general, organ repair played a leading role on the 4th day after modeling.

Children with Cryptorchidism Complicated by Testicular Torsion: A Case Series.

OBJECTIVE: To investigate the clinical features, diagnosis, treatment and prognosis of children with cryptorchidism complicated by testicular torsion. METHODS: The clinical data of 6 children with cryptorchidism complicated by testicular torsion admitted to our hospital from December 2000 to December 2016 were analyzed retrospectively. RESULTS: All 6 children were diagnosed with cryptorchidism by surgery, their age was from 12 days up to 11 years, and the average time between onset of symptoms and diagnosis was 20.5 h. Torsion testis was located in the groin area and the rate of left to right was about 2:1. Twist was 600° on average. All children were admitted because of the inconsolable cry, abdominal pain, and the swelling of the groin. Three patients underwent orchidectomy, while the other 3 patients underwent detorsion and cryptorchidopexy. Color Doppler ultrasound examination showed normal testes at 6 months after operation. Only 1 case was diagnosed with cryptorchidism after birth. CONCLUSIONS: Cryptorchidism is an emergency in pediatric urology and often leads to a low testicular salvage rate, especially in infants, due to lack of knowledge, delayed diagnosis, and late treatment. Neonatal genital examination is important for the early diagnosis and management of cryptorchidism.

[Serum metabolomics of preterm and full-term infants based on gas chromatography-mass spectrometry].

OBJECTIVE: To study the features of serum metabolites in preterm infants based on gas chromatography-mass spectrometry (GC-MS), and to find differentially expressed metabolites in the serum of preterm infants. METHODS: Serum samples were collected from 19 preterm infants and 20 full-term infants before feeding. GC-MS was used to measure metabolic profiles, and the metabolic features of 397 serum metabolites in preterm infants were analyzed. RESULTS: There was a significant difference in serum metabolic features between the preterm and full-term infants before feeding. There were significant differences between the full-term and preterm infants in the levels of metabolites such as O-phosphonothreonine, digicitrin, tannic acid, and fructose-1,6-diphosphate (P<0.01), suggesting that the above differentially expressed metabolites were highly differentiated between the preterm and full-term infants. Most differentially expressed metabolites were involved in the metabolic pathways such as ABC transporters, ß-alanine and pyrimidines and were correlated with some clinical parameters (albumin and total bilirubin) (P<0.05). CONCLUSIONS: There is a significant difference in serum metabolites between preterm and full-term infants before feeding. Metabolomics plays an important role in improving metabolic disorders and exploring metabolism-related diseases in preterm infants.

Altered Levels of Zinc and N-methyl-D-aspartic Acid Receptor Underlying Multiple Organ Dysfunctions After Severe Trauma.

BACKGROUND: Severe trauma can cause secondary multiple organ dysfunction syndrome (MODS) and death. Oxidative stress and/or excitatory neurotoxicity are considered as the final common pathway in nerve cell injuries. Zinc is the cofactor of the redox enzyme, and the effect of the excitatory neurotoxicity is related to N-methyl-D-aspartic acid receptor (NMDAR). MATERIAL AND METHODS: We investigated the levels of zinc and brainstem NMDAR in a rabbit model of severe trauma. Zinc and serum biochemical profiles were determined. Immunohistochemistry was used to detect brainstem N-methyl-D-aspartic acid receptor 1 (NR1), N-methyl-D-aspartic acid receptor 2A (NR2A), and N-methyl-D-aspartic acid receptor 2B (NR2B) expression. RESULTS: Brain and brainstem Zn levels increased at 12 h, but serum Zn decreased dramatically after the trauma. NR1 in the brainstem dorsal regions increased at 6 h after injury and then decreased. NR2A in the dorsal regions decreased to a plateau at 12 h after trauma. The levels of NR2B were lowest in the death group in the brainstem. Serum zinc was positively correlated with NR2A and 2B and negatively correlated with zinc in the brain. Correlations were also found between the brainstem NR2A and that of the dorsal brainstem, as well as between brainstem NR2A and changes in NR2B. There was a negative correlation between zinc and NR2A. CONCLUSIONS: Severe trauma led to an acute reduction of zinc enhancing oxidative stress and the changes of NMDAR causing the neurotoxicity of the nerve cells. This may be a mechanism for the occurrence of MODS or death after trauma.

[Iron and ferritin changes in multiple organs failure after trauma].

OBJECTIVE: To investigate the changes of iron content in serum and liver, ferritin content in serum, percentage of myeloperoxidase (MPO) positive granulocyte in rabbits after different serious trauma and to explore the relationship between these changes and multiple organ failure (MOF). METHODS: Rabbit trauma models were established. Iron content in serum and liver, ferritin content in serum and the percentage of MPO positive granulocyte were measured at different time after trauma. RESULTS: After trauma, iron content in serum decreased sharply in early period (12-36h) and increased gradually to normal level in mild traumatic group after 60 h. Iron content in serum remained lower level in severe traumatic and death group 60 h after trauma. Iron content in liver obviously increased in death group. The changes of ferritin content in serum in mild traumatic were not obvious. Ferritin contents in serum in severe injury group and death group were slightly higher in early period and decreased in later period. The percentage of MPO positive granulocyte increased in early period after trauma. The percentage began to decrease 6 d after trauma and returned to normal level in mild traumatic group. The percentage obviously was significantly lower than normal levels in severe traumatic group and death group 6 d after trauma. Some rabbits died 60 h-6 d after severe trauma, and the pathological changes in the other organs were consistent with MOF. CONCLUSION: Trauma can cause the serum iron, ferritin levels and percentage of MPO positive granulocyte changes. Severe trauma can cause uncompensated changes of these indicators, which could be the main mechanisms of MOF and death.

Gas chromatography-mass spectrometry based serum metabolic analysis for premature infants and the relationship with necrotizing enterocolitis: a cross-sectional study.

BACKGROUND: Preterm birth and feeding are the most important pathogenic factors of neonatal necrotizing enterocolitis (NEC). Metabonomic has been widely used in the diagnosis and treatment of other diseases, but there is no research on the related diseases of premature infants. Compared with full-term infants, the metabolism of preterm infants has its own specificity, so it can easily lead to NEC and other digestive tract inflammatory diseases. Metabonomic may be applied to the diagnosis of preterm related diseases, such as NEC. METHODS: The model was established with premature infant serum samples from 19 premature infants in our hospital, which was compared with the full-term infant control group. Serum was analyzed by gas chromatography-mass spectrometry (GC-MS), coupled with the analysis of serum metabolic characteristics. The variable important in projection, P value and Pearson correlation coefficient of samples were analyzed by using SIMCA, SPSS and other multivariate statistical analysis software. RESULTS: Compared to the term infants, premature infants had significantly higher levels of luteolin, and lower levels of xylose, O-succinyl-L-homoserine and lauric acid in the serum. There were some correlations among several different metabolites and clinically related indices (albumin, total bilirubin) for premature birth related diseases. CONCLUSIONS: There are metabolic alterations in the serum of premature infants, which make contribution to the diagnosis of NEC.

[A morphological study with serial histological slices on the normal and abnormal gubernacula in newborn male mice].

OBJECTIVE: To explore the feasibility of serial slices microscopic histological investigation for the elaborate evaluation of reproductive system malformations. METHODS: Newborn male mice prenatally exposed to different doses of subcutaneously given diethylstilbestrol (DES) from gestational day 9 to 17 were treated by fixing parts of the abdomen in situ and setting them to transected serial slices. All the slices were stained, studied under the microscope and serially recorded by software. The gubernaculum was morphologically analyzed and its location and size were measured. RESULTS: Morphologically, the gubernaculum could be identified clearly, its structure inhomogeneous from proximal to distal and dissymmetric from right to left. The environmental estrogen produced different effects on the morphology of the gubernaculum in different parts and most obviously affected its length. CONCLUSION: Prenatal exposure to environmental estrogen has evident and general effects on the gubernacular development of newborn male mice. The morphological study with serial histological slices gives a precise and systematic evaluation of genital malformations.

Changes of serum trace elements in early stage trauma and its correlation with injury severity score.

BACKGROUND: Severe trauma can cause secondary multiple organ dysfunction syndrome and death. The absolute and relative concentrations of trace elements in both critical care and conventional treatment, which can lead to acute trace element deficiency, constitute an important mechanism of multiple organ dysfunction syndrome (MODS)/multiple organ failure (MOF). METHODS: We investigated the changes in serum Cu, Zn, and Fe in early stage trauma of patients with the high injury severity score (ISS) and correlated the change in trace elements with ISS. Blood samples were collected within an hour of admittance and the patients were scored according to ISS. We collected clinical data records and ISS score values, and determined serum Fe, Zn, and Cu by inductively coupled plasma mass spectrometry. RESULTS: Compared with the control group, the serum Zn and Fe values of trauma patients were decreased. There was no significant difference in serum Cu between the patients and the control group. In the trauma group, the serum Zn and Fe were lower than that of the minor injury group, and the difference of Cu concentration was not statistically significant. CONCLUSIONS: Serum Zn and Fe levels in patients with multiple trauma fractures were significantly different than those in the normal group, suggesting that Zn and Fe need to be monitored in the early stage of trauma.

Peutz-Jeghers syndrome with intermittent upper intestinal obstruction: A case report and review of the literature.

RATIONALE: Peutz-Jeghers syndrome (PJS) is an autosomal dominant genetic syndrome characterized by a unique type of gastrointestinal hamartomatous polyp associated with oral and anal mucocutaneous pigmentations. Peutz-Jeghers polyps occur most numerously in the small intestine but frequently in the colon and stomach, only a few cases have been reported in the duodenum. PATIENT CONCERN: A further family history survey discovered 10 out of 14 members of the family (in 4 generations) had mucocutaneous pigmentations, but many of them were living in rural areas where they had no access to specialized medical services, so none were checked with endoscopy for polyps of hamartoma. DIAGNOSES: We report the case of a boy patient with mucocutaneous pigmentations over the lips, and a history of recurrent bouts of vomit and anemia over the preceding two years, no abdominal pain and mass. An upper gastrointestinal endoscopy revealed some small polyps in the stomach and multiple sessile polyps in the second part of the duodenum, but colonoscopy exam did not reveal any lesion. INTERVENTIONS: A double polypectomy and duodenum segmentary resection with end-to-end anastomosis was performed. Histopathology of the resected duodenum polyps indicated it to be a typical hamartomatous polyp. OUTCOMES: The child was under regular follow-up and recovered well. LESSONS: In this case, the patient was characteristic with pigmentations on his lips and intermittent upper intestinal obstruction (due to mass duodenal polyps), there are no definitive guidelines for the treatment to duodenal PJS hamartomatous polyp, each case requires tailor-made management.

Diagnosis and treatment of pediatric benign pneumoperitoneum: A case report series of 9 patients.

INTRODUCTION: Benign pneumoperitoneum (BPPT) is defined as asymptomatic free intraabdominal air or as pneumoperitoneum without peritonitis. Symptomatic free air requires surgical anagement, but management of asymptomatic pneumoperitoneum is controversial. In this study, we investigate the diagnosis and treatment of BPPT in children. CLINICAL FINDINGS: The clinical data of 9 pediatric patients with BPPT who were admitted to our hospital from January 2000 to January 2015 were retrospectively analyzed to summarize the diagnosis and treatment. Overall, 9 cases were included with 8 males and 1 female, aged from 4 days to 4 years. Among them there were 6 newborns (including 1 premature infant). Patients were all admitted to hospital with the major clinical symptom of abdominal distension, including 2 cases accompanied by tachypnea, 2 cases with vomiting, 1 case with diarrhea, and 2 cases with fever. No previous constipation or obstructive defecation existed. Six newborns had meconium defecation within 24 hours after birth. Physical examination revealed all patients with relaxed abdominal wall except 1 patient with abdominal distension had slight muscle stiffness and hyperactive bowel sounds. Abdominal X-ray suggested free air under the diaphragm in all cases. INTERVENTIONS/OUTCOMES: All patients except for one case of laparotomy were conservatively treated and cured with fasting, infection prevention, rehydration, abdominocentesis, and close observation. Nine cases of patients were all discharged with no death occurrence. After discharge follow-up of 7 months to 6 years was conducted. There was no recurrence of similar symptoms, and children were in good growth and development. CONCLUSION: The diagnosis of BPPT mainly relies on clinical symptoms in patient, careful abdominal examination, abdominal X-ray combined with abdominocentesis, and the exclusion of gastrointestinal perforation for confirmation. Conservative treatment can cure the disease. Attention should be paid to distinguish with surgical pneumoperitoneum to avoid unnecessary surgical exploration.

Expression of glucocorticoid receptor isoforms in cutaneous hemangiomas and vascular malformations.

BACKGROUND: Hemangiomas are the most common tumors in children. Some hemangiomas may require intervention because of their location, size, behavior, or potential for important complications. Pharmacological therapy with glucocorticoids is the mainstay treatment, but there is no consensus on therapeutic regimens or candidate selection, therapeutic efficacy varies, and the mechanism mediating the beneficial effects of glucocorticoids remains unclear. This study was performed to investigate the expression patterns of the glucocorticoid receptor (GR) and its alpha isoform (GRalpha) in cutaneous hemangiomas and vascular malformations. METHODS: SP immunohistochemical technique was used to examine the expression of GR(e-20) (GR) and GR(p-20) (GRalpha) on vascular endothelial cells in 80 specimens that included 33 proliferating hemangiomas, 32 involuting hemangiomas, 7 vascular malformations as well as 8 normal skin tissues, all obtained from infants and children. GR and GRalpha expression in prepared tissue slides were examined using automated computer-assisted microscopic analysis. Mean gray scale values were compared among the various tumor types. RESULTS: The mean gray scale values of GR were 127.0 +/- 6.4 and 121.4 +/- 6.6 in hemangiomas and vascular malformations respectively, but this difference was not statistically significant (P = 0.104). However, these values were all markedly higher than that of normal skin, which was only 108.6 +/- 6.8 (P = 0.001 and P = 0.000 for comparison with hemangiomas and vascular malformations respectively). The gray scale of GR in proliferation and involuting hemangiomas were 127.9 +/- 4.8 and 126.0 +/- 5.8 respectively, but this difference was not significant (P = 0.146). However, GRalpha expression in hemangiomas, vascular malformations and normal skin declined gradually in stepwise fashion (127.3 +/- 5.4, 120.4 +/- 6.1 and 109.9 +/- 5.3 respectively; P < 0.001). GRalpha expression was higher in proliferating hemangiomas than in involuting hemangiomas (127.2 +/- 6.3 and 122.5 +/- 6.3; P = 0.004). CONCLUSIONS: GR and GRalpha are strongly expressed in hemangiomas and vascular malformations. The expression of GRalpha is closely related to the phase of the hemangioma. Determination of GR and GRalpha may be a positive significance to understand the information of hemangiomas and vascular malformations and may further help determining proper strategies of steroid therapy for hemangiomas and vascular malformations.

Blood zinc, iron, and copper levels in critically ill neonates.

The aim of this study is to explore the prognostic value of blood zinc, iron, and copper levels in critically ill neonates by comparing blood metal levels with the score for neonatal acute physiology (SNAP). Forty-six neonates (26 boys, 20 girls; ages ranging from 10 min to 23 days) who had been admitted to the neonatal intensive care unit of hospital and who were critically ill according to SNAP were included. Another 15 neonates (12 boys, 8 girls; ages ranging from 30 min to 24 days) who were brought to the hospital for a health checkup were included as controls. Clinical data, time in the intensive care unit, prognosis, and SNAP for critically ill neonates were recorded. Blood Cu, Zn, and Fe values were measured by inductively coupled plasma atomic emission spectrophotometry. Ill neonates were divided into extremely critical (SNAP ≥ 10) and critical groups (1 ≤ SNAP < 9). Zn levels were lower in patients than in controls (p <0.05). Cu levels did not differ between patients and controls (p >0.05). Fe levels were not significantly between the critical and control groups (p >0.05). In ill neonates, blood Zn and Fe concentrations in the extremely critical group were lower than in the critical group (p <0.05). Serious illness in neonates may lead to decreased Zn and Fe blood concentrations. Zn and Fe supplements may be beneficial for critically ill children.

Computer-aided three-dimensional reconstruction of main vessels in hemangiomas.

This study aimed to investigate three-dimensional (3-D) morphological features of the main vessel architecture of human hemangioma. Serial sections of specimens from three cases of children hemangioma were stained with hematoxylin and eosin (HE) to visualize the vessels. Serial images were taken and processed with computer-assisted 3-D reconstruction. Partial 3-D structure reconstruction of vessel morphology in hemangioma revealed strange distribution and branching, which were different from normal vessels of the human skin. The 3-5 microvascular was most common in hemangioma. We observed respective characteristics of three cases: 1 case showed uniform artery vein distribution accompanied by running trend; 1 case showed main artery distribution and less vein distribution, and there were many blood sinus in the shallow surface close to the skin; another case showed vein distribution in the middle of antrum. In conclusion, digital vascular model of 3-D structure of main vessel hemangioma provides a new way for the diagnosis and treatment of hemangioma of children.

Prognostic value of blood zinc, iron, and copper levels in critically ill children with pediatric risk of mortality score III.

We aimed to explore the association of blood Zn, Fe, and Cu concentrations and changes in the pediatric risk of mortality (PRISM) score in critically ill children, to predict prognosis. We included 31 children (22 boys and 9 girls, 1 month to 5 years old), who had been admitted to the intensive care unit of our hospital and who were critically ill according to PRISM score of III. Another 20 children (12 boys, 8 girls, 3 months to 5 years old) who were brought to the hospital for a health checkup were included as controls. We recorded clinical data, time in the intensive care unit, prognosis, and PRISM III score for critically ill children. Blood Cu, Zn, and Fe values were measured by inductively coupled plasma atomic emission spectrophotometry. Zn and Fe levels were significantly lower in patients than in controls (all p < 0.05). Cu levels differed between patients and controls, but not significantly (p > 0.05). In ill children, blood Zn and Fe concentrations were inversely correlated with PRISM III score (Zn: r = -0.36; Fe: r = -0.50, both p < 0.05), with no significant correlation of blood Cu level and PRISM III score (r = -0.13, p > 0.05). Serious illness in children may lead to decreased Zn and Fe blood concentrations. Zn and Fe supplements may be beneficial for critically ill children.

Altered levels of trace elements in acute lung injury after severe trauma.

Trace element (TE) supplementation can reduce the incidence of multiple organ failure after severe trauma. The lung plays a main role in post-injury multiple organ failure. In the present study, the relationship between TEs and acute lung injury (ALI) post-injury was investigated in a rabbit model of severe trauma with an injury severity score of 27. New Zealand white rabbits were randomly assigned to trauma-control, trauma-TE groups, and a control group. During days 1-5 post-trauma, each rabbit in the trauma-TE group received 0.1 ml multi-TE compound intraperitoneally to give a daily dose of 32.50 mg/kg of Zn, 6.35 mg/kg of Cu, 1.38 mg/kg of Mn, and 0.16 mg/kg of Se. Concentrations of blood and lung selenium (Se), copper (Cu), zinc (Zn), and manganese (Mn) were measured at 6 and 24 h, as well as 3, 6, 9, and 14 days after trauma. Levels of glutathione peroxidase (GPx), total superoxide dismutase (SOD), Cu/Zn superoxide dismutase (Cu/Zn-SOD), and malondialdehyde (MDA) in serum and lung tissue and the level of intercellular adhesion molecular-1 (ICAM-1) in serum were detected simultaneously. In addition, the lung coefficient (LC) and the lung permeation index (LPI) were measured. Serum and lung Zn, Se, and Mn levels decreased dramatically by 6 h after trauma in both experimental groups. Cu showed no significant changes after trauma. The serum and lung GPx and SOD levels in the experimental group decreased significantly on days 1 and 3, respectively. Serum and lung MDA began to increase on day 3 in the trauma group but increased less after TE supplementation. Serum ICAM-1 peaked on day 6 in the experimental group. LC and LPI increased gradually post-trauma, peaking on days 6 and 9, respectively. In conclusion, an acute lung injury causes declines of the levels of TEs in serum and lung which can be significantly prevented by TE supplementation and which can also mitigate some of the morphological and biomechanical changes in ALI.