Objective To investigate the blood levels of amino acids in children aged 0-15 year, with an attempt to provide evidence for evaluating amino acid status and diagnosing metabolic diseases of amino acid.Methods The blood levels of eleven amino acids in 1900 children aged 0-15 years were determined by tandem mass spectrometry (MS/MS). Results The blood levels of leucine & isoleucine, valine, phenylalanine tyrosine,glycine, proline, ornithine, and alanine gradually decreased after birth, reaching the lowest levels at the ages of 4-6 months, and then gradually increased, reaching the normal range at the ages of 7 months-1 year. The blood levels of alanine and glycine reach the second peaks on the ages of 9 years in girls and on 11 years in boys. The blood levels of methionine and arginine were lowest in the first week of age, became highest in 1-3 months, decreased to the normal ranges after 4-6 months, and kept the level afterwards. The ratios between prosomatic amino acid and productive amino acid, between ornithine and arginine, between citrulline and arginine, and between ornithin and citrulline were highest in the first week of age and decreased to normal values after 3 to 12 months. The concentrations of amino acids in group of 7 months-15 years were significantly different from the group of 1 day-1 month and group of 2-6 months (P ＜0. 05 or P ＜0. 01 ). The concentrations of amino acids were significantly higher in females than in males in the group of 1 day-1 month and in all age groups (P ＜0. 05 or P ＜0. 01 ) . Conclusions The concentrations and profiles of amino acids change remarkably during the first year of age. Age should be carefully considered when evaluating the nutritional status of amino acid and diagnosing metabolic diseases of amino acids.

Objective　To develop the newborn screening and treatment for congenital hypothyroidism (CH) in order to prevent the children from growth and mental retardation and to improve the population quality in China. Methods　TSH value in the dried-blood spot speciments on the filter papter was measured by time-resolved fluorescence immunoassay (Tr-FIA). Clinical analysis for the CH patients was performed. Results　Among 315　　472 neonates screened for CH, 66 (47 overt and 19 subclinical) cases of CH were confirmed and the incidence of CH was 1/4　　780. Among the 47 typical cases, 2 infants were recognized with transient hypothyroidism, 2 infants were not reevaluated because of below 2 year old, 1 infant died, 1 case gave up treatment, the rest 41 infants were permanent CH. Among the 19 atypical cases, 7 cases were recognized with permanent CH, 4 cases with transient disease, 5 cases with hyperthyrotropinemia, 3 cases were not evaluated because of below 2 years old at that time. The patients followed in the clinic are all currently with normal growth and intellectual development. Conclusion　Our center is the first labolatory to screen CH with Tr-FIA in China. Tr-FIA is an ideal non-radioactive technology. The typical patients as well as atypical cases should be all treated using L-thyroxine. It is essential to arrange a trial treatment for confirming the diagnostic results from screening and to decide whether they need a lifetime treatment.

Objective To provide methodological references for laboratories to carry out newborn screening for disorders of amino acid metabolism,we compared the difference and distribution of ten amino acids including alanine (Ala),arginine (Arg),citrulline (Cit),glycine (Gly),leucine (Leu),methionine (Met),ornithine (Orn),phenylalanine (Phe),tyrosine (Tyr),and valine (Val) from newborn dried blood spots specimen using derivatization or non-derivatization as sample preparation methods.Method It is a comparative research study.A total of 4135 newborn screening dried blood spot samples for inborn errors of metabolism were collected from January to June,2012.All specimens came from neonatal screening center of shanghai children's hospital.Samples were prepared by two different techniques,the corresponding kits and the procedures were used as follows:(1) Simultaneous detection of 100 dried blood spot specimens using two methods respectively to compare the paired difference of each amino acid.(2) 2000 cases of normal newborn specimens were detected respectively to obtain the normal distribution of ten neonatal amino acids.(3) 35 specimens from patients previously diagnosed positively as inborn errors of metabolism were simultaneously detected with 7 amino acids to verify the consistency of two techniques in clinical judgment.Results The amino acid levels of normal newborns analyzed by one-sample.kolmogorovSmirnov test (Z value ranged from 1.997 to 6.229) showed a skewed distribution (P ＜ 0.01).Except for Leu and Tyr,non-derivatization techniqueshowed a lower concentration than derivatization technique,and the CVs of nine amino acids were ＜ 10％ except for Met (the CV of Met was 47.8％),and the average CV is 7.8％.Except for Met,Phe and Tyr,the levels of other 7 amino acids measured by two techniques showed no significant difference (P ＞ 0.05).According to 0.5th to 99.5th percentiles,the normal reference range for derivatization method were greater than on-derivatization method,and the average value was 25.3％.After clinic judgment,the results of the abnormal indicators of children with true metabolism disorders showed no statistically significant between two methods (P ＞ 0.05),the detection rate was 100％.Conclusions There was a slight difference between derivatization and the non-derivatization techniques in detecting multiple amino acids.The results of the abnormal indicators of amino acid metabolism disordersshow no statically significant difference between the two methods,and no difference in clinical judgment.Both methods can be used in detecting amino acid metabolism disorders in newborn screening.

Objective To summarize the acylcarnitine profile in children with malnutrition,with an attempt to distinguish it from those of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency,multiple acylCoA dehydrogenase (MAD) deficiency,or glutaric aciduria type Ⅱ (GA Ⅱ).Methods Thirteen pediatric patients with malnutrition and 214 children of the same age but without malnutrition,which was set as the control group,were included in this study.The blood samples were collected at admission,and the concentration of carnitine and acylcarnitines were measured in bloodspots by tandem mass spectrometry using samples nnderivatized.Results The concentrations of acylcarnitines which were involved in fatty acid oxidation,including octadecanoyl (C18) to acetyl (C2) acylcarnitines and ketonic acylcarnitines,were higher in malnutrition group than in the control group.Particularly,the concentration of decanoyl acylcarnitine (C10) in the malnutrition group was (0.203 ±0.105) μmol/L,which was out of the normal rang (0-0.200 μmol/L),was significantly higher than that [(0.054 ±0.030) μmol/L] in the control group (P ＜0.001).There was no significant difference in the concentrations of acylcarnitines [e.g.propionyl (C3),isovaleryl (C5),3-hydroxy-isovaleryl (C5OH),and glutaryl (C5DC) acylcamitines] involved in amino acid decomposition between the malnutrition and control groups.Conclusions The concentrations of acylcarnitines related to fatty acid oxidation elevate in children with malnutrition.In particular,the medium-chain acylcarnitines C10 is out of the normal range,which can be used to differentiate malnutrition from MCAD and MAD.

Objective:To evaluate the seven existing vaginal birth after cesarean (VBAC)screening tools and to identify additional factors that may predict VBAC or failed trial of labor in China.Methods:In the study,53 patients with 1 previous cesarean delivery who then delivered between January 1,2007 and Novenber 31,2014 were recruited.The average age of the patients was (32.1 ±3.5 )years,the average gestational age was (38.0 ±2.3)weeks.There was no significant difference of the successful group and the failed group in the maternal /neonatal mortality and morbidity,also in the incidence of the postpartum hemorrhage and the postpartum infection.The probability of VBAC was calculated for each participant using 7 prediction models created by Weinstein,Flamm,Grobman,Gonen,Troyer,Smith and Torri.The data were analyzed using t test,rank-sum test,and receiver operating curve analysis. Results:44 trial of labor patients had a vaginal birth after cesarean delivery,and the successful rate was 83%.The scores between the successful group and the failed group had significant difference when eva-luated by Weinstein and Grobman scoring models only.After recalculating the successful rate of VBAC in different score levels according to the references,there was significant difference between the rates of dif-ferent score levels when evaluated by the Weinstein model.The successful rates of different score levels were higher compared to the references (<50%)when evaluated by the Troyer (70%),Gonen (60%),Torri (85.7%)models.The area under the receiver operating characteristic curve of Weinstein prediction model (0.746)and Flamm prediction model (0.723)were more than 0.7,and there was no significant difference between the seven models.Conclusion:Among the seven scoring models,the Weinstein model is more applicable to the population of our country,but a new model more applying to Chinese women still needs to be created.

Objective: To study the dietary habits and serum leptin levels of obese children and its possible association to lipids and glucose metabolism, serum lipids and C peptide (CP) in children with different body weight. Methods: 52 obese children were inquired about their favorite foods and food intake. 145 healthy children (85 boys, 60 girls), aged 6-13, with a wide range of body mass index (BMI) 14 to 30 kg/m 2, were measured for fasting serum leptin, CP, glucose, lipids (TG, TC, LDL C,HDL C), height and weight. The children were classified into three groups by BMI, i.e. normal weight group (A group), overweight and slight obese group (B group). moderate and severe obese group (C group). Results: 65% of obese children liked meat best. 38.46% of them ate most in supper. The serum leptin, CP, TG in obese children were higher than that in normal children. Leptin concentration of girls were higher than that in boys in A and B groups (P

Objective To develop a method of detecting dry blood spot (DBS) samples collected in the filter paper by using Surface Enhanced Raman Spectroscopy (SERSp,also known as Raman molecular fingerprint spectrum) technology.The possibility and reliability of applying the technology to the newborn screening of congenital hypothyroidism (CH) was investigated.Methods The case-control study was used.The results of CH-hTSH screening test based on the DBS samples collected in the filter paper,and the clinical diagnosis were all provided by neonatal screening center of Shanghai Children's Hospital.The results of SERSp analysis were provided by Dalian University of Technology.Six positive and six negative samples,which were confirmed respectively by CH-hTSH screening using time resolved fluoroimmunoassay (TRFIA) were chosen.With the spectrum lines of 725 cm-1 as internal standard,the SERSp from the aqueous solution of these samples were analyzed.The specific spectrum line analysis,the principal component analysis (PCA) and the scatter diagram of PCA were used to compare the results of TSH test.Results With the spectrum lines of 725 cm-1 as internal standard,the CH specific spectrum lines of 1373 cm-1 and 1400 cm-1 were discovered.They showed the most significant difference of the peak height between the positive and negative samples of CH,while TSH specific spectrum lines of 785 cm-1,827 cm-1 and 853 cm-1 failed to distinguish positive from negative samples with distinct peak height.The consistency between the results of CH-Raman Screening and CH-hTSH Screening was 91.7％ (11/12),which was proven by PCA,the scatter diagram of PCA and the specific spectrum line analysis.The sensitivity of CH-Raman Screening was 100％ and the specificity was 83.3％.Conclusion There is a new CH-Raman Screening method using the SERSp analysis,which was proven to be a promising technology in the newborn screening of congenital hypothyroidism with the DBS in the filter paper.

Objective To study the prognosis of gastroesophageal reflux disease ( GERD) in children, and explore the factors which impacts on the prognosis of GERD. Methods One hundred and thirteen children with GERD were enrolled on the basis of positive result of 24-hour pH-monitoring between January 2007 and November 2011. The number of patients who were followed up was 87,and the parents of children were contacted with the telephone. The prognosis was evaluated by comparing the degree of patients′symptom relief,and the cumulative symptom relief rate was calculated by Kaplan-meier product limit method. The univariate Log-rank test and the COX proportional hazardmodel multivariate analysis were applied to detect the factors impacting on the prognosis,including age,gender,the regularity of treatment,reflux index,and Boix-Ochoa standard score,with esophageal hiatal hernia or without,receiving surgical treatment or not,the diet and lifestyle improved or not,receiving anti-acid treatment or not,as well as with allergies his-tory or without. Results At last,76 out of 87 children had symptom relieved. Survival curve showed the cumulative symptom relief rate at different time points,the median cumulative symptom relief rate reached 6 months,the final relief rate was close to 90. 0%,and the continuous treatment time was 44 months. The study showed that 14. 9% (13/87 ca-ses) of children′s growth and development were affected and the life and learning in 16. 1% (14/87 cases) of children were impacted. Age (P=0. 012,Wald=6. 376) and the regularity of treatment (P=0. 000,Wald=13. 059) were the risk factors in the prognosis of GERD. Conclusions Age and the treatment regularity were the factors in the prognosis. The children aged more than 1-year old have poor prognosis compared with those less than 1-year old,and the irregular treatment is the risk factor in the prognosis.

BACKGROUND:Experiments have demonstrated that biological membranes can be usedtorecon struct thetendon she athandin hibit exogenou shealing of thetendon.Therefore,the semembrane sprovide a good bed for tendon gliding and reduce tendon adhesion.
OBJECTIVE:To compare the effectsof acelular amniotic membrane and medical membraneagainst tendon adhesion during the repair oftendon sheath defects.
METHODS:ToesIIIfrom the bipeds of 66 leghorns were chosen to prepare tendon injury and tendon sheath defect models, which were randomly divided into three groups (n=22 per group). Amnion group were repaired with acelular amniotic membrane, medical membrane group with absorbable membrane, and control group had no treatment on tendon sheath defects. Gross, histological and biomechanical tests of each group were performed at 2, 4, 8, 12 weeks after surgery.
RESULTS AND CONCLUSION:At 12 weeks after surgery, in the amniotic membrane and medical membrane groups, the tendon sheath formed completely, and the tendon healed well, with no adhesion, but in the control group, there was serious tendon adhesion. At 8 weeks after surgery, the number of synovial cells in the false sheath was highest in the amniotic membrane group sequentially followed by the medical membrane group and control group. In the amniotic membrane group, the rough endoplasmic reticulum expanded highly and secreted exuberantly in the matrix, while in the control group, the synovial cells presented with messy arrangement, and expanded vacuoles in the matrix were weaker than those in the other two groups. At 12 weeks after surgery, fibroblasts were arrayedtidily in layerwith dense structure in the medical membrane and amniotic membrane groups;but in the control group, fibroblasts were distributed disorderly with loose structure. Tendon sliding distance and total flexor toe angle in the amniotic membrane and medical filmgroups were significantly larger than those in the control group (P < 0.05),butthere was no significant difference between the medical membrane and amniotic membrane groups. Additionally, the maximum tensile fracture strength had no significant difference among three groups at 12 weeks after surgery. These results indicate that both amniotic membrane and medical membrane can markedlyprotect the tendon from exogenous healing and adhesion.

BACKGROUND:To improve local microenvironment and reduce local scars is conducive to peripheral nerve regeneration that promotes nerve function recovery.OBJECTIVE:To evaluate the effect of fresh amniotic membrane on the regeneration of tinjured peripheral nerve.METHODS:Sixty healthy Sprague-Dawley rats were randomly divided into three groups (n=20 per group) after constructing a model of sciatic nerve injury of the unilateral leg. In group A, the nerve was wrapped with fresh human amnion at the anastomosis end after the repair of nerve. In group B, the nerve was wrapped with biofilm at the anastomosis end after the repair of nerve. In group C, no treatment was conducted after the repair of nerve (blank control). The effects were evaluated by anatomical observation, light microscope observation, immunohistochemical detection (2, 4, 8, 12 weeks after surgery), transmission electron microscope observation, axon imaging analysis, action potential detection, and sciatic nerve function index (4, 8, 12 weeks after surgery).RESULTS AND CONCLUSION: (1) Gross observation. The amniotic membrane and biofilm were absorbed partialy at postoperative 2 weeks, mostly at postoperative 4 weeks and completely at postoperative 8 weeks. In the groups A and B, the nerve was adhered slightly and loosely to the surrounding tissues, with a fair range of motion. In the group C, the nerve was tightly adhered to the surrounding tissues, with a poor range of motion. (2) Observation under light microscope. The nerve regeneration was better in the groups A and B than group C at 2, 4, 8, 12 weeks postoperatively. (3) Observation under electron microscope. Regenerated nerve fibers were rarely seen and lamelar structures were unclear in the three groups at 4 weeks postoperatively. Then, increased regenerated nerve fibers, thickened myelin sheath, clear lamelar structure and enlarged axon diameter were found in the groups A and B compared with the group C at 8 and 12 weeks postoperatively. (4) Immunohistochemical detection. The expression and distribution of S-100 protein in the groups A and B were better than those in the group C. (5) Axon image analysis. Groups A and B were superior to the group C in the diameter of myelinated nerve fibers, thickness of myelin sheath and number of regenerated nerve fibers. There was a significant difference by statistical analysis (P < 0.05). (6) Electrophysiological examination. Shorter latency period, higher amplitude and faster nerve conduction velocities were observed in the groups A and B compared with the group C (P < 0.05). (7) The sciatic function index. The sciatic function index in group A or B was significantly higher than that in group C (P < 0.05). To conclude, the human amniotic membrane can reduce adhesion between the damaged nerve and surrounding tissues, and prevent scarring at the anastomosis end. In addition, it promotes the regeneration of nerve fibers, increase axon diameter and myelin sheath thickness, and ease inflammatory and immune responses at the neural incision.