Serum Bile Acid Levels in Preoperative and Postoperative Patients with Congenital Biliary Atresia

To elucidate the bile acid metabolism in the preoperative and postoperative stages of congenital biliary atresia (CBA), the unconjugated and conjugated bile acid levels in sera were measured by high performance liquid chromatography (HPLC). The results showed that the mean total serum bile acid (TSBA) level in the preoperative cases of CBA was higher, 122.1±39.0 n?mol/1 (1SD), and was about 12 times higher than the level in the age-matched normal controls (9.9±6.0). Even in the non-icteric patients several years after operation, the mean TSBA level was still much higher (15.2±9.6) than that of normal controls (5.7±3.1). The mean ratio of cholic acid to chenodeoxycholic acid (CA/CDCA) in the preoperative and postoperative cases of CBA was less than 1.0. The mean ratio of glycine-conjugated bile acids to taurine-conjugated bile acids (G/T) in the preoperative cases of CBA was the lowest (1.9±1.1). In the clinically good cases with sufficient bile flow after operation, G/T ratio was the highest (9.3±6.5). In the normal controls and the patients with preoperative and postoperative CBA, the main bile acids were glycocholic acid, taurocholic acid, glycochenodeoxycholic acid and taurochenodeoxycholic acid. These conjugated bile acids comprised more than 85% of the total in amount.     

Studies on the Enterohepatic Circulation of Bile Acids in Infancy and Childhood

To elucidate the enterohepatic circulation of bile acids in hepatobiliary disorders, the present author measured the fasting cholic acid levels in serum and followed up the changes of the levels after MCT milk administration. The subjects were 17 cases of neonatal hepatitis, 24 cases of congenital biliary atresia (CBA), 19 cases of other hepatobiliary disorders and 117 normal children. The serum cholic acid levels in the neonatal period were significantly high, which suggested a physiological cholestasis in neonates which gradually decreased with age. The mean level in CBA was rather higher than that in neonatal hepatitis but showed overlap of the levels. The patterns of changes of serum cholic acid levels in MCT milk test were classified into 6 types which were respectively characteristic of each disorders according to varied disturbance of the enterohepatic circulation of bile acids. This MCT milk test may be useful in making a differential diagnosis of various hepatobiliary disorders, especially neonatal hepatitis and CBA.     

The Screening of Patients with Extrahepatic Biliary Atresia by Measuring Total Bile Acids Absoredin Dried Blood Spots

The efficency was assessed of the screening for extrahepatic biliary atresia by measuring total bile acids absorbed in dried blood spots at around 5 days of age. When the cut-off level was 54.0 μmol/l, the sensitivity, the specificty and the efficiency of the screening test were 87.9%, 93.9% and 93.9% respectively. The case-finding rate was 0.009% in this type of screening which was three times as many as an early case-finding rate by population screening. These results suggest that teh efficiency of this screening is acceptable.     

IMMUNOREACTIVE TRYPSIN SCREENING FOR CYSTIC FIBROSIS

ABSTRACT. Immunoreactive cationic trypsin (irCT) was measured in 22 cystic fibrosis (CF) and 132 control infants. IrCT was analysed with radioimmunoassay of dried blood samples collected for PKU screening around the 5th day of life and stored on filter paper. The mean ± 1 SD level of irCT for the control infants was 42±19 μg/l. Sixteen of the 22 CF children had an irCT level above 100 μg/l (mean + 3 SD) while 6 had a level at or below this cut-off limit. A specificity of 99%, which gives a sensitivity of 73%, and an approximative noise: signal ratio of 30: 1, suggests that the irCT test may be unsatisfactory as a neonatal screening method for CF.     

THE PLASMA LEVELS OF 25-HYDROXYVITAMIN D IN PATIENTS WITH VARIOUS LIVER DISEASES AND THE RESPONSE OF 25-HYDROXYVITAMIN D TO VITAMIN D TREATMENT

ABSTRACT. The mean plasma levels of 25-hydroxyvitamin D (25-OH-D) were measured before and after the administration of 2000 units of daily oral vitamin D₂ for a period of 2 weeks in 9 normal infants and children, 7 infants with neonatal hepatitis and persistent neonatal hepatitis, and 4 infants with congenital biliary atresia. The mean plasma level of 25-OH-D increased significantly from 19.5±3.7 (S.E.) ng/ml to 34.0±6.8 (S.E.) ng/ml after administration of vitamin D₂ in controls ( p <0.05). The mean plasma level of 25-OH-D also increased from 8.0±2.1 (S.E.) ng/ml to 22.1±2.6 (S.E.) ng/ml after vitamin D treatment in hepatitis group ( p <0.05). In patients with congenital biliary atresia, vitamin D treatment did not affect the plasma levels of 25-OH-D.     

URINARY MONOHYDROXY BILE ACIDS IN YOUNG INFANTS WITH OBSTRUCTIVE JAUNDICE

ABSTRACT. Using an aluminum oxide column, we fractionated and quantitatively determined urinary monohydroxy bile acids in young infants. For comparison purposes, monohydroxy bile acids were also measured in urine from older children and adults with obstructive jaundice. Lithocholic acid was not found in any specimens of the young infants examined, while 3β-hydroxy-5-cholenoic acid was detected in all. In the biliary atresia group, 3β-hydroxy-5-cholenoic acid excreted was 0.45 ± 0.28 μmol per day ( n = 7), and in the neonatal hepatitis group, 0.48 ± 0.44 μmol per day ( n = 9). The mean rate of 3β-hydroxy-5-cholenoic acid to total urinary bile acids in the biliary atresia group was 2.1%, and 1.3% in the neonatal hepatitis group. In the older children and adults with obstructive jaundice ( n =6), 3β-hydroxy-5-cholenoic acid was excreted at a mean rate of 3.9% of total urinary bile acids, ranging from 0.63 to 14.81 mol per day. The excretion rate of 3β-hydroxy-5-cholenoic acid was related to that of chenodeoxycholic acid ( p <0.05) in infants, while it was related to that of both chenodeoxycholic acid ( p <0.01) and cholic acid ( p <0.05) in older children and adults.     

Radioimmunoassay of Thyroxine in Dried Blood Spots

Sadler, W. A., Lynskey, Clare P., and Legge, M. (1978). Aust. Paediatr. J., 14, 154–182. Radioimmunoassay of thyroxine in dried blood spots. A rapid precise radioimmunoassay for thyroxine (T4) in dried blood spots on filter paper has been developed, and appears entirely suitable for use as a mass screening procedure for the early diagnosis of neonatal hypothyroidism. A systematic study of sampling factors which may influence levels of measured T4 suggests that non-uniformity of sample preparation is the single greatest error source In a blood spot assay. The assay reaction mixture consists of a 4.5 mm diameter disc punched from the blood spot and 8-anilino-1-naphthalene sulphonic acid, labelled T4 and T4 antiserum in 1.1 ml barbitone buffer. Following incubations for one hour at 37°C and overnight at 4°C, bound and free hormone are separated using dextran coated charcoal. Assay standards consist of a 2:3 mixture of a standardized serum and washed red cells. In 558 Infants screened to date T4 levels are not normally distributed, being skewed towards the lower end, with a median value of 179 nmol/l. One Infant with congenital hypothyroidism has been found.     

血清内毒素水平在胆道闭锁患儿术后的意义

目的探讨血清内毒素(ET)、透明质酸(HA)与胆道闭锁(BA)患儿术后肝损害、肝纤维化的关系。方法以行肝门空肠吻合(Kasai)术后的BA患儿为BA组(n=38),正常体检儿童(n=12)为对照组;BA患儿根据其胆红素水平再分成黄疸组(n=14)和无黄疸组(n=24)。由自动生化仪测肝功能,鲎三肽基质显色法测ET水平、放射免疫法测HA水平。结果 BA组患儿存在肝损害,血清ET、HA水平均较正常对照儿童高(P<0.05);BA组患儿中,黄疸患儿的肝损害较无黄疸者重,血清ET、HA水平也较无黄疸者高,差异均有统计学意义(P<0.05);血清ET水平与血清总胆红素、谷草转氨酶及HA水平呈正相关(P<0.05)。结论血清ET水平与术后BA患儿肝损害和肝纤维化密切相关,可作为术后判断BA患儿预后的有用指标。     

Smoking during Pregnancy–Effects on the Fetal Thyroid Function

ABSTRACT. Infants delivered at term by mothers smoking at least 10 cigarettes daily during pregnancy ( n =46) were found to be growth retarded compared to infants of non-smoking mothers ( n =49), birthweights 3445pM385 (SD) g and 3667pM392 g respectively ( p <0.05) in the two groups. Cord serum thyrotropin (TSH) was significantly decreased (8.2±4.(1 U/l vs. 10.3±4.9 U/l) and free thyroxine index (FT₄I)/TSH ratio significantly increased (18.8±9.0 vs. 14.4±7.6) ( p <0.05) in the smoking group compared to infants of non-smokers. Cord serum thyroxine (T₄) and FT₄I were higher in the smoking group (149.0±22.4 nmol/1 and 125.5± 14.9 respectively) compared to infants of non-smoking mothers (140.6±21.6 nmol/1 and 120.0±16.5 respectively), with borderline statistical significance (0.05< p <0.10). The results indicate that infants of smoking mothers may have a hyperfunction of the thyroid gland at birth compared to infants of non-smokers, with a negative feed-back on TSH production from the pituitary gland. Increased metabolic rate and oxygen consumption caused by fetal thyroid hyperfunction may be pathogenetic factors for the fetal growth retardation caused by maternal smoking.     

Inter-relationship between serum concentrations of glucose, glucagon and insulin during the first two days of life in healthy newborns

The relationship between serum concentrations of glucose, insulin and glucagon during the first two days of life was studied in healthy newborns. The first capillary blood sample was obtained at 3–15 h of age (median 6h; day 0) and a second sample approximately 24 h later (day 1). Serum glucose concentrations in the first sample averaged 2.1 ± 0.07mmol/l (mean ± SEM; n = 60) and were positively correlated with postnatal age ( p < 0.01). Serum glucagon concentrations in the first sample averaged 570 ± 32pg/ml and were inversely correlated with glucose concentrations ( p < 0.0001). At the second sampling, serum glucose concentrations had increased to 2.9 ± 0.07mmol/l ( p < 0.001; n = 57) and serum glucagon concentrations had decreased to 403 ± 22pg/ml ( p < 0.001). Serum insulin concentrations were 11.7 f 0.3 μU/ml and 10.2 ± 0.3 μU/ml at the two samplings and did not correlate with serum glucose concentrations. The relationship of serum glucose and hormone concentrations to maternal and infant characteristics was studied by stepwise regression analysis. Serum glucose concentration on day 0 was positively correlated with postnatal age ( p < 0.01) and birth weight ( p 0.05) but inversely correlated with duration of labour ( p < 0.05). Serum glucose concentration on day 1 was positively correlated with birth weight ( p < 0.0001) and inversely correlated with maternal prep-pregnancy weight ( p < 0.05). Similar analyses of serum hormone concentrations did not demonstrate any relationships with maternal or infant characteristics. It is suggested that glucagon secretion is part of the counter-regulation against hypoglycaemia in healthy newborns and that neonatal energy stores, as indicated by birth weight, influence the ability to increase circulating glucose concentrations in response to counter-regulatory hormones.     

Short-term effects of blood transfusion on blood volume and resting peripheral blood flow in preterm infants

The effects of blood transfusion on cardiac output and blood pressure are variable, but resting peripheral blood flow (RPBF) may be a sensitive indicator of changes in blood volume. The purpose of this investigation was to study the effects of red cell transfusion on blood volume (Evans blue), blood pressure, RPBF in the leg (strain-gauge plethysmography) and blood viscosity (cone-plate viscometer) in preterm infants during the first week after birth. Fourteen infants with mean ± SD birth weight of 1658 ± 429 g, gestational age 33 ± 3 weeks and postnatal age 64 ± 40 h received 18 ±4 ml/kg of packed red cells (red cells 11±2 ml/kg, plasma 7± 1 ml/kg) because their hematocrit was less than 0.45 l/l. Mean blood volume before transfusion was 88±15 ml/kg. The increase in blood volume (9 ±4 ml/kg) measured 4 to 6 h after transfusion was smaller than the transfused volume (18 ± 4 ml/kg), due to a shift of plasma to the extravascular space. The plasma shift increased with increasing pretransfusion blood volume ( r = 0.70; p = 0.007). Red cell transfusion caused an increase in RPBF by 25% ( p <0.01), whereas systolic blood pressure (BP) increased by only 12%. Peripheral resistance (R = BP/RPBF) decreased by 9% (p<0.01). Blood viscosity (±) increased by 21'% ( p <0.001) and vascular hindrance (R/±) decreased by 24% ( p < 0.001), indicating vasodilatation of limb arteries. The increase in RPBF and the decrease in hindrance were particularly pronounced in infants with high pretransfusion blood volume. We conclude that the increase in blood volume after transfusion is not proportional to the transfused volume and that RPBF increases more than systolic blood pressure with increasing blood volume. The increase in RPBF can be explained by vasodilatation of limb arteries and by increased blood pressure.     

脐血可溶性瘦素受体与胎儿生长发育关系探讨

目的：瘦素是肥胖基因的蛋白产物,除参与调节机体能量代谢外,与胎儿的生长发育密切相关,但其作用机制尚不清楚。本研究旨在探讨脐血可溶性瘦素受体与胎儿生长发育的关系及其可能机制。方法：67例足月新生儿根据出生体重分为小于胎龄儿(SGA)组23例,适于胎龄儿(AGA)组44例。采用ELISA法测定脐血和母血中瘦素及可溶性瘦素受体水平,并采用体脂含量估测新生儿营养状态。结果：①脐血可溶性瘦素受体水平与脐血瘦素水平及新生儿出生体重、体脂含量呈负相关(r分别为-0.405,-0.366,-0.356,P均0.05)。③SGA组脐血可溶性瘦素受体明显高于AGA组［(18.24±6.02) ng/ml vs (13.80±4.37) ng/ml］,P<0.01;而SGA组的瘦素含量低于AGA组［(6.79±4.59) ng/ml vs (16.30±11.62) ng/ml］,P<0.01。④脐血可溶性瘦素受体水平男性高于女性［(16.89±4.37) ng/ml vs (13.95±5.29) ng/ml］,P<0.05;而脐血瘦素水平则是男性低于女性［(10.28±8.28) ng/ml vs (15.70±12.11) ng/ml］,P<0.05。结论：可溶性瘦素受体可能通过对血清中游离瘦素水平的调节实现对胎儿生长发育的调控作用。同时测定瘦素和可溶性瘦素受体可能更有利于进一步了解瘦素的病理生理作用机制。     

Interleukin-1 receptor antagonist in the fetomaternal compartment

Interleukin-1 (IL-1) is a major mediator in infections and inflammation. Interleukin-1 receptor antagonist (IL-1ra) opposes the actions of IL-1. IL-1ra is present in exceptionally high concentrations in third trimester amniotic fluid. We studied IL-1ra in amniotic fluid, fetal serum and newborn urine. The concentrations of IL-1ra in amniotic fluid at mid-trimester and at 25-41 gestational weeks were 6.6 ± 0.5ng/ml ( n = 30) and 100 ± 4ng/ml ( n = 202), respectively. At mid-trimester, amniotic fluid IL-1ra was not dependent on fetal gender, whereas during the third trimester IL-1ra was higher in female- than in male-bearing gestations. Urine of normal term newborns during the first day of life contained a very high concentration of IL-1ra (125 ± 16ng/ml, n = 50). Urinary concentration in female newborns was significantly higher than that in male newborns (202 ± 19ng/ml, n = 25 versus 49 ± 14ng/ml, n = 25). IL-1ra concentration in fetal serum at 22-36 gestational weeks was 0.50 ± 0.07ng/ml ( n = 31) and at term 1.5 ± 0.3ng/ml ( n = 17). Serum concentrations were not gender-dependent. The gender differences in IL-1ra concentrations may in part explain the lower susceptibility of female fetuses to infection.     

Bone γ-Carboxyglutamic Acid Containing Protein in the Perinatal Period

ABSTRACT. We measured bone γ-carboxyglutamic acid-containing protein (BGP), calcium (Ca), phosphorus (P), and alkaline phosphatase (Al-P) in paired maternal and cord sera, and urinary γ-carboxyglutamic acid (γ-GIa) in neonates. The circulating BGP was 41.21±2.47 ng/ml and 7.44±0.87 ng/ml in the cord (n=15) and the maternal (n=14) sera, respectively. The urinary γ-GIa in the neonates was 147.68 ± 10.75 μ.mol/g creatinine (n=15). The cord serum BGP was significantly higher than the normal adult level. The maternal serum BGP was at the same level as in other adults. It is conceivable that the fetus may produce BGP during gestation, as the cord serum BGP level was significantly higher than the maternal level and there was no correlation between the cord and maternal serum BGP concentrations. The reason for the elevated circulating BGP level in the cord serum is not known, but increased bone turnover may be a factor. The cord serum BGP may include not only carboxylated but also non-γ-carboxylated BGP because of fetal vitamin K deficiency.     

Increased lipid peroxidation in children with autoimmune diseases

To examine the role of oxidative damage in children and adolescents with autoimmune diseases, we compared blood serum levels of the lipid peroxidation (LPO) products 4-hydroxynonenal (HNE) and malondialdehyde (MDA) in 22 children with systemic lupus erythematosus (SLE), 13 children with focal type of scleroderma, and 21 healthy controls. In order to study the influence of disease activity in SLE on serum LPO product levels, the SLE group was divided into one group with active disease ( n = 11) and one group with non-active disease ( n = 11) according to SLEDAI-score, 15.1 and 1.8, respectively. SLE patients with active SLE (146 ± 14nmol/l, median 145nmol/l) have significantly higher HNE levels compared to controls (61 ± 10nmol/l, median 52nmol/l), whereas the MDA serum levels are similar to those of the control group, 1.94 ± 0.18μmol/l (median: 2.02μmol/l) and 1.58 ± 0.11 μmol/1 (median: 1.52 μmol/l), respectively. Children with SCL had HNE and MDA levels similar to the control group.     

Hyaluronic acid: Additional biochemical marker in the diagnosis of biliary atresia

BACKGROUND: The purpose of the present paper was to evaluate the value of biochemical markers, including conventional liver function tests, gamma-glutamyl transferase (GGT), and hyaluronic acid (HA), in the diagnosis of neonatal cholestasis. METHODS: Infants with neonatal jaundice were consecutively enrolled during 1 year period. The patients were diagnosed as having biliary atresia (BA) if there was either bile ductular proliferation in the portal tracts, atretic common bile duct/gallbladder, or evidence of bile duct obstruction demonstrated by liver pathology or intraoperative cholangiography, respectively. Serum HA was measured using an enzyme-linked immunosorbent assay-based test. RESULTS: A total of 25 patients diagnosed as having BA (n = 10), neonatal hepatitis (NH; n = 9), choledochal cyst (n = 3) and parenteral nutrition-induced cholestasis (n = 3), were studied. The age at diagnosis was not significantly different between groups. Only GGT and HA were significantly elevated in the patients with BA when compared to NH (P = 0.02, P = 0.03, respectively). In BA, the median value of serum HA was 514 ng/mL (range 19-4476 ng/mL), compared to 50 ng/mL (range 19-315 ng/mL) in NH. Additionally, the serum HA level was much higher in children with choledochal cyst. CONCLUSION: HA could be considered as a complementary biochemical marker for evaluating infants with prolonged jaundice.     

The role of fever on cerebrospinal fluid glucose concentration of children with and without convulsions

In febrile convulsions glucose concentrations are known to increase both in the blood and cerebrospinal fluid (CSF). The reason behind this increase is, however, incompletely understood. We have studied the effects of convulsion and fever on the CSF and blood glucose concentrations in four different groups of children: febrile and non-febrile children, with and without convulsions. The concentration of glucose in the CSF was significantly higher in febrile children with (4.4 ± 0.1 mmol/1, mean ± SEM n = 35, p < 0.01, ANOVA, Duncan's test) and without convulsions (3.9 ± 0.2mmol/1, n = 22, p < 0.05) than in non-febrile, non-convulsive children (3.3 ±0.1 mmol/1, n = 21). In non-febrile convulsive children, the CSF glucose concentration was 3.7 ± 0.2mmol/l (n = 10). Both fever and seizures increased the CSF glucose levels (p < 0.0001 and p = 0.028, respectively, analysis of covariance). There was a linear correlation between the body temperature and concentration of glucose in the CSF (r = 0.454, p < 0.0001, n = 88, Pearson's correlation analysis). The changes in blood glucose concentrations between the groups parallelled those found in the CSF. Our results show that hyperglycaemia and an increase in the CSF glucose concentration in febrile convulsions is not explained just by a stress reaction, evoked by the seizure, as has been hypothesized earlier, but by the influence of increased body temperature as well.     

Influence of Mode of Delivery on Fetal Pituitary-Thyroid Axis

We measured the cord serum levels of thyroid-stimulating hormone (TSH), thyroxine (T4) and triiodothyronine (T3) in 922 neonates delivered by mothers who had no thyroid disorders. The T4 and T3 levels increased progressively from the 27th to the 40th week of gestation. However, the TSH levels varied widely and had no correlation with gestational age, because they were affected by the mode of delivery. The mean cord serum TSH level in neonates delivered by vacuum extraction was 16.3 ±10.0 µUml (n = 30), which was significantly higher than the level following normal vaginal delivery (9.5 ± 6.0 µ U/ml, n = 622) (p < 0.005). The mean cord serum TSH level following elective caesarean section was 6.5 ± 3.1 µ U/ml (n = 79), and this was significantly lower than after normal vaginal delivery (p< 0.005). TSH levels in high-risk neonates were significantly higher than in neonates without risk factors. A significant positive correlation was found between the duration of the second stage of labor and the cord serum TSH level (r = 0.45, n = 412, p<0.01). However, there was no correlation between the cord serum TSH level and the congenital hypothyroidism screening TSH level (r = 0.01, n = 468). We conclude that the cord serum TSH level reflects delivery stress and that an elevated level does not influence the congenital hypothyroidism screening TSH test in which blood is obtained at five days of life.     

新生儿血清瘦素水平与生长发育关系研究

目的：探讨新生儿血清瘦素与生长发育的关系。方法：采用放射免疫法检测80例新生儿静脉血和脐血瘦素水平,其中66例足月儿分为大于胎龄儿(LGA)组18例,适于胎龄儿(AGA)组32例,小于胎龄儿(SGA)组16例。采用Rohrer’s指数=出生体重(g)×100/身长(cm)~3估测新生儿营养状态。结果：早产儿血清瘦素水平明显低于足月儿[(0.66±1.03)ng/ml vs(3.59±2.16)ng/ml],P<0.01;足月儿中AGA血清瘦素水平[(3.06±0.96)ng/ml]明显低于LGA[(4.03±2.22)ng/ml],而高于SGA[(1.13±1.98)ng/ml];足月新生儿血清瘦素水平与Rohrer’s指数、新生儿体重、胎龄呈显著正相关(r=0.61,0.68,0.62,P均<0.01)。结论：新生儿体内瘦素是反映新生儿的发育和营养状态的有用指标。[中国当代儿科杂志,2003,5(1):29-30]     

Determination of free thyroxine (FT4) for the detection of connatal hypothyroidism within the scope of neonatal screening

Using the reagents of a commercially available test kit (Henning, Berlin) for the determination of free thyroxin (FT4) in serum we developed a radio immuno assay procedure to measure FT4 in whole dried blood on filter paper. The inter assay coefficient of variation was 19.1%, 10.7%, and 11.0% for the hypo-, normo-or hyperthyroid range for whole blood on filter paper. The corresponding within assay values were 7.0, 10.3, and 5.3% respectively. In 15,793 samples of dried blood on filter paper which were collected on the 5th or 6th day of life for the screening of inborn errors of metabolism or hypothyroidism FT4 was measured in addition to our routine determination of thyroid stimulating hormone (TSH). The mean concentration of FT4 was 20 to 22 pg/ml. 97,8% of all results were found between 10 and 35 pg/ml. Connatal hypothyroidism was confirmed clinically in 5 children showing FT4-values of 3.5, less than 2.0, less than 2.0, 3.4, and 6.2 pg/ml. The corresponding TSH-values were greater than 224, greater than 224, 99, 203, and 129 microU/ml. To make sure that a sufficient amount of blood had been eluted from the filter paper we measured the concentration of hemoglobin as an additional parameter. Low FT4 together with low hemoglobin concentrations were obtained in 18 samples indicating that a mismeasurement of FT4 might have occurred. We suggest that the determination of TSH in doublicates, which is the routine screening procedure in West Germany might be replaced by a combined determination of FT4 and TSH.