Newborn screening for fatty acid oxidation disorders / 中国实用儿科杂志

()of recessive hereditary diseases caused by the dysfunction of enzymes required for fatty acids to enter mitochondria or fatty acid beta-oxidation,including carnitine transport disorders and fatty acid beta-oxidation disorders. Clinical symptoms are non-specific,involving multiple organs,such as liver,myocardium,skeletal muscle,brain and kidney. Most FAOD patients diagnosed by newborn screening have no clinical symptoms or mild symptoms through early intervention management,but they are prone to acute onset or even sudden death under stress conditions such as hunger and exercise. Long-term follow-up and management can effectively reduce the mortality and morbidity rate of FAOD.

The under-five mortality rate and the causes of death in Zhejiang Province between 2000 and 2009 / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the under-five mortality rate and the causes of death in Zhejiang Province between 2000 and 2009 in order to provide a basis for reducing the mortality rate in the region.</p><p><b>METHODS</b>By stratified random cluster sampling, all the children under 5 years old from 30 sampling areas of Zhejiang Province between 2000 and 2009 were enrolled. The under-five mortality rate and the leading causes of death were investigated by descriptive analysis and Chi-square test.</p><p><b>RESULTS</b>The under-five mortality rate demonstrated a decreased trend in Zhejiang Province between 2000 and 2009, with the under-five mortality rate of 14.83‰ in 2000 compared to 9.49‰ in 2009. In 2009, the under-five mortality rate in rural regions was significantly higher than that in urban regions (9.14‰ vs 6.50‰; P<0.01). Compared with the resident population, there were an increased under-five mortality rate in floating population (12.12‰ vs 6.42‰; P<0.01). Preterm delivery/low birth weight was the top death cause in children under 5 years old. The top three causes of death in infants less than 1 year old were preterm delivery/low birth weight, congenital heart disease and birth asphyxia compared to drowning, traffic accident and falling in children aged 1-4 years.</p><p><b>CONCLUSIONS</b>There are differences in the under-five mortality rate between rural and urban children as well as between the floating and resident population in 2009. The leading causes of death in different age groups are different. It is essential to reduce the mortality rate by preventing preterm delivery, low birth weight and congenital malformations to infants and preventing accidental injuries to children aged 1-4 years.</p>

Screening for neonatal inborn errors of metabolism by electrospray ionization-tandem mass spectrometry and follow-up / 中华儿科杂志

<p><b>OBJECTIVE</b>To determine the impact of expanded newborn screening using tandem mass spectrometry (MS/MS) on the overall detection rate of inborn errors of metabolism in Zhejiang province and to assess the outcome of the patients who were diagnosed.</p><p><b>METHOD</b>Blood spots were collected between days 3 and 6 of life from the newborns. All samples were subjected to MS/MS analysis using Waters Quattro API. Confirmation tests included amino acid analysis, urinary organic acids by GC-MS, routine blood analysis, biochemistry, blood gas analysis, blood glucose and ammonia tests, blood homocysteine, lactate and pyruvate tests, urine acetone tests, biotin and biotin enzyme profile and DNA analysis. Standard treatment protocol was given to the patients. Protein restricted diet, special powdered formula and medicines recommended for the patients with amino acidemias. Protein restricted diet and L-carnitine, folic acid and Vitamin B12 supplementation were given for the patients with organic acidemia. L-carnitine was given to the patients with primary carnitine deficiency. The overall epidemiology, prognosis, follow-up of the screening program were also investigated in the neonates.</p><p><b>RESULT</b>A total of 129 415 neonates were investigated for 26 inborn errors of metabolism during the period. Twenty-three newborns were confirmed as having inborn errors of metabolism, including 13 with amino acidemias, 6 with organic acidemias and 4 with fatty acid oxidation disorders. The prevalence was 1:5626. Positive predictive value was 2.10%, specificity was 99.72% and sensitivity 100%. Seventeen children remain asymptomatic during the follow-up. Five patients had motor and mental developmental delay. One patient presented metabolic disorders during the follow-up. No death occurred in this series of patients.</p><p><b>CONCLUSION</b>This strategy represents a valuable preventive medicine approach by enabling diagnosis and treatment before the onset of symptoms.</p>

Behavioral development in adolescent rats of perinatal hypothyroidism and its relations to androgen receptor gene expression in hippocampus / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the behavioral development in adolescent rats of perinatal hypothyroidism and its relation to androgen receptor (AR) gene expression in the hippocampus.</p><p><b>METHODS</b>Perinatal hypothyroidism was induced by gavages 50 mg/d of propylthiouracil solution in 48 dams starting at embryonic day 15 through the lactation period. Twenty-four pups (M:F=1) of perinatal hypothyroidism were injected intraperitoneally with 2 microg T(4)/100 g BW daily from the day of birth to the age of 21 days (treatment group); 24 pups (M:F=1) without treatment were designated as hypothyroidism group. And 24 normal pups (M:F=1) served as the control group. The effects of perinatal hypothyroidism on the abilities to learn and retain memory traces and on behavior were observed in rats of both sexes at 60 days. Experiments were performed using models of conditioned "open" field test and passive avoidance reflexes. Hippocampus samples were collected and AR mRNA was detected by competitive RT-PCR.</p><p><b>RESULT</b>Perinatal hypothyroidism caused an increase of crossing number and decrease of rearing and defecation in both sexes. In treatment groups, only the crossing number in male didn't reach the normal level (P >0.05). In passive avoidance test, hypothyroidism groups showed more mistakes in both sexes and shorter latencies in males, the females performed better than males (P <0.01). The treatment groups performed significantly better than the age-matched hypothyroidism groups and reached the normal level (P >0.05). AR mRNA levels in hippocampus of hypothyroid group were lower than those of the controls in males, and the levels in treatment groups were significantly higher in comparison with the hypothyroidism groups (P <0.01). There were no significant differences among the three female groups (P >0.05). In male group, there was negative correlation between the number of crossing and AR mRNA in the hippocampus (r=-0.537, P=0.001), negative correlation between the number of mistake and AR mRNA (r=-0.532, P=0.001), and positive correlation between the latency and AR mRNA (r=0.564, P=0.000).</p><p><b>CONCLUSION</b>Perinatal hypothyroidism results in hyperactivity and anti-anxiety effects on adolescent rats, the sex difference is depleted, and also causes learning and memory impairment but the degree of influence higher in male than female. The decreased level of AR mRNA expression in hippocampus contributes to the change of behavioral ability in adolescent male.</p>

Effects of hypothyroidism on apoptosis and the expression of Bcl-2 and Bax gene in the neonatal rat hippocampus neurons / 中华儿科杂志

<p><b>OBJECTIVE</b>During the critical period of brain development, insufficiency of thyroid hormone results in severe mental retardation and learning deficit. This study was designed to investigate the effects of hypothyroidism on apoptosis and the expression of Bcl-2 and Bax gene in the developing rat hippocampus neurons and to explore the mechanism of brain development regulated by thyroid hormone.</p><p><b>METHOD</b>Hypothyroidism was induced by administration of propylthiouracil (PTU, 50 mg/d) solution to the dams from gestational day 15 by gavage. Pups from both hypothyroid and control groups were harvested at postnatal day 1 (P1), P5, P10 and P15, respectively. Blood samples were collected at the time of death for the determination of thyroid hormone. Serum free tri-iodothyronine (FT(3)) and free thyroxine (FT(4)) were measured by using chemoluminescence. Hippocampus collected from the control and hypothyroid pups were examined under light and transmissional electron microscopy. Measurement of DNA fragmentation was carried out by agarose gel electrophoresis. The expression of Bcl-2 and Bax protein in the developing rat hippocampus neurons was performed by Western blotting.</p><p><b>RESULTS</b>Significantly lower circulating FT(4) and FT(3) levels confirmed the hypothyroid status of the experimental pups. The shrunken and contracted degenerations increased in hippocampus neurons of hypothyroid pups under light microscopy. Enhanced apoptotic cells were found in hippocampus neurons of hypothyroid pups under transmission electron microscopy, especially at P10 and P15. Extensive DNA fragmentation was seen throughout development in hippocampus of hypothyroid pups, but not in the euthyroid controls except for basal level at P10. The expression of Bcl-2 in the hippocampus neurons of hypothyroid pups was significantly lower than that of euthyroid controls at all stages of development (P1: 1.95 +/- 0.27 vs. 2.59 +/- 0.19, P < 0.05, P5: 1.86 +/- 0.24 vs. 2.47 +/- 0.17, P < 0.05, P10: 1.29 +/- 0.22 vs. 1.86 +/- 0.28, P < 0.05 and P15: 1.21 +/- 0.27 vs. 2.18 +/- 0.17, P < 0.01, respectively). The relative amount of expression varied significantly with age in the control pups. The level of Bcl-2 was high in hippocampus neurons of euthyroid at P1, P5, and decreased significantly at P10, and showed a trend of recovery at P15. Similar age-related variation in the expression of Bcl-2 gene was observed in the hypothyroid group at P1, P5 and P10, but the level was maintained low at P15. The expression of Bax in the hippocampus neurons of hypothyroid pups was significantly higher than that of control pups at all stages of development (P1: 1.69 +/- 0.14 vs. 1.24 +/- 0.23, P < 0.05, P5: 1.78 +/- 0.16 vs. 1.29 +/- 0.17, P < 0.05, P10: 1.92 +/- 0.18 vs. 1.45 +/- 0.14, P < 0.05 and P15: 1.86 +/- 0.14 vs. 1.51 +/- 0.12, P < 0.05, respectively). The ratio of Bcl-2/Bax in hippocampus neurons of hypothyroid pups was lower than that of age-matched controls (P1: 1.16 +/- 0.17 vs. 2.12 +/- 0.35, P < 0.05, P5: 1.05 +/- 0.16 vs. 1.94 +/- 0.36, P < 0.05, P10: 0.68 +/- 0.17 vs. 1.29 +/- 0.16, P < 0.05 and P15: 0.67 +/- 0.19 vs. 1.45 +/- 0.22, P < 0.01, respectively).</p><p><b>CONCLUSION</b>Thyroid hormone significantly prevents apoptosis of hippocampus neurons. Congenital hypothyroidism increases not only the extent but also the duration of apoptosis by down-regulation of the anti-apoptotic gene Bcl-2 and maintaining a high level of the pro-apoptotic gene Bax.</p>

Expression of peripheral blood neutrophil CD64 in neonatal septicemia / 中华儿科杂志

<p><b>OBJECTIVE</b>Neonatal septicemia is a common and severe infection, which often results in death. Early diagnosis and treatment of neonatal septicemia may help decrease neonatal mortality. Recently, many studies sought to explore the possibility of early diagnosis of this disease. The high affinity Fcgamma-receptor I (CD(64)) was purposefully chosen as a potential marker for identifying neonatal septicemia. The present study was designed to evaluate neutrophil CD(64) level for early diagnosis of neonatal septicemia.</p><p><b>METHODS</b>Eighty-nine suspected neonatal septicemia cases were recruited into the study. Five non-specific indices, i.e., C-reactive protein (CRP), micro-erythrocyte sedimentation rate (mESR), white blood cell count, platelet count and the ratio of immature neutrophil count to total neutrophil count were measured for each patient. The patients were divided into septicemia group (n = 39) and non-septisemic infection group (n = 50) according to the diagnostic criteria for neonatal septicemia. Nineteen hospitalized neonates with non-infectious diseases were enrolled as controls (n = 19). The levels of peripheral blood neutrophil CD(64) were measured by using flow cytometry. The positive rate, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of CD(64) were calculated.</p><p><b>RESULTS</b>The levels of peripheral blood neutrophil CD(64) in septicemia patients were (75.6 +/- 8.9)%, which were significantly higher than those of non-septisemic infection group (29.1 +/- 6.2)% and control group (5.1 +/- 1.1)% (P < 0.05), respectively. There were no significant differences in the levels of CD(64) expression between the patients with Gram-negative (79.5 +/- 3.5)% and Gram- positive (76.4 +/- 5.0)% (P > 0.05) bacterial infection. The levels of CD(64) of the cases with septicemia significantly decreased at day 10 of treatment with antibiotics. The detection of CD(64) (cutoff value > 30%) for suspected septicemia showed high sensitivity (97.4%), specificity (84.0%), PPV (82.6%), and NPV (97.6%). The positive rate of CD(64) detection (62.9%) was much higher than that of the blood culture test (19.1%) and that of the five nonspecific indices (29.2%, P < 0.05, respectively).</p><p><b>CONCLUSION</b>The expression of CD(64) increased in neonatal septicemia cases. The measurement of cell surface expression of CD(64) on neutrophils may be helpful to early diagnosis, evaluation of severity of infection and observation of therapeutic effects for neonatal septicemia.</p>

Expression of androgen receptor gene in cerebral cortex and hippocampus of rats with perinatal hypothyroidism / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of perinatal thyroid hormone deficiency on the expression of androgen receptor (AR) mRNA in cerebral cortex and hippocampus of rats.</p><p><b>METHODS</b>Perinatal hypothyroidism was induced by the administration of propylthiouracil (PTU) solution to the dams by gavage (50 mg/d) beginning at embryonic d15 throughout the lactational period. In the T(4) injected group hypothyroid rats were injected intraperitoneally with levothroxine (L-T(4)) 2 microg/100 g BW daily, starting from the day of birth. Cerebral cortex and hippocampus specimen were collected from controls,hypothyroid and T(4)-injected hypothyroid rats on postnatal d1, 5, 10, 15 and 20. Quantification of ARmRNA in cerebral cortex and hippocampus was performed with competitive RT-PCR using internal and external standardization.</p><p><b>RESULT</b>Age-related increasing ARmRNA levels were observed in neonatal rats, and those in male animals were significantly higher. AR expression was higher in the hippocampus than in the cerebral cortex. ARmRNA levels in the hypothyroid pups were lower than those in age-matched controls. The mRNA levels in the T(4)-injected hypothyroid pups were significantly higher compared with the age-matched hypothyroid pups, but in hippocampus ARmRNA expression did not reach normal levels in male rats at d10 and d20, in female at d15 and d20.</p><p><b>CONCLUSION</b>The expression of ARmRNA decreases in brain of rats with perinatal hypothyroidism. Treatment with thyroid hormone can recover ARmRNA expression in cerebral cortex, but not in hippocampus.</p>

Mechanism for apoptosis of hippocampus neuron induced by hypothyroidism in perinatal rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the mechanism for the apoptosis of hippocampus neuron induced by hypothyroidism in perinatal rats.</p><p><b>METHODS</b>Hypothyroidism was induced by administration of propylthiouracil (PTU, 50 mg/d) solution to the dams from gestational day 15 by gavage. Pups from both hypothyroid and control groups were harvested at 1, 5, 10 and 15d, respectively. Blood samples were collected at the time of death for the determination of thyroid hormone. Serum free triiodothyronine (FT(3)) and free thyroxine (FT(4)) were measured by chemoluminescence. Hippocampus specimens were collected from the control and hypothyroid pups.Mitochondia was examined under transmission electron microscopy. Translocation of apoptogenic molecules (Bax, cytochrome C and AIF) and activation of caspase-3 were analyzed by Western Blotting.</p><p><b>RESULT</b>Significantly low circulating FT(3) and FT(4) levels confirmed the hypothyroid status of the experimental pups. Electron microscopy showed that altered morphology of mitochondria significantly increased under hypothyroid conditions. The expression of Bax in the cytosol of hypothyroid pups was higher than that of control pups at all stages of development (P<0.05),and significantly higher in mitochondria (P<0.001). The expression of cytochrome c in the cytosol of hypothyroid pups was significantly higher than that of control pups at all stages of development (1,10 and 15 d:P<0.05, 5d: P<0.001), and lower in mitochondria (P<0.05). The expression of AIF in the cytosol of hypothyroid pups was higher than that of control pups at all stages of development (P<0.001), and significantly lower in mitochondria (1, 5d: P<0.001, 10, 15 d: P<0.01). he expression of caspase-3 P20 in the cytosol of hypothyroid pups was significantly higher as compared with that of the age-matched controls (1, 15d: P<0.01, 5,1 0 d: P<0.001).</p><p><b>CONCLUSION</b>The intrinsic death pathway in mitochondria may be one of the mechanisms with which hypothyroid induces apoptosis of hippocampus neuron in developing rats.</p>

On the behaviour,learning and memory in rats with perinatal hypothyroidism / 中华内分泌代谢杂志

The results of open-field test,step-through passive avoidance task and radial-arm maze experiment showed that changes of locomotor activity,anxiety-related behaviour and ability of learning and memory were associated with the increased apoptosis of neurons in hippocampus in rats with perinatal hypothyroidism.