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 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.

Background High myopia is one of the primary factors of visual impairment,and its prevention and management are researching hot topics.Corneal curvature (CC) measures the steepness of the cornea which is an important parameter leading to myopia.Genome-wide association study (GWAS) showed that several genes are associated with CC in Asian populations.However,the association of corneal curvature-related genes with high myopia is unclear up to now.Objective This study was to investigate the association between single nucleotide polymorphism (SNP) in the rs74225573 (mechanistic target of rapamycin [MTOR]),rs60078183 (cytidine/uridine monophosphate kinase 1 [CMPK1]),rs1800813 (platelet derived growth factor receptor alpha [PDGFRA]),rs11204213 (retinol binding protein 3 [RBP3]) and high myopia in Chinese Han population.Methods A prospective cohort study was performed.Four hundreds and eighty-three patients with high myopia were collected in Sichuan Provincial People's Hospital from February 2012 to August 2013,with the diopter (-10.84±4.69)D in the right eyes and (-10.35±4.67)D in the left eyes or ocular axial length of (28.15±2.27)mm in the right eyes and (27.72±2.51)mm in the left eyes.Five hundreds and nineteen normal volunteers matched in age and gender were included in the same period as controls,and all the subjects were Chinese Han people without genetic relationship.The periphery blood of 4 ml was obtained for the DNA extraction from each subject under the written informed consent.The primers of rs74225573,rs60078183,rs1800813 and rs1 1204213 were designed based on the information of NCBI website.The four SNPs were amplified by real-time PCR and genotyped by SNaPshot method.Results All the genotype frequencies of these four SNPs were in Hardy-Weinberg equilibrium (HWE).There are no significant differences in minor allele frequency (MAF) distribution of rs74225573,rs60078183 and rs11204213 between high myopia group and normal control group (rs74225573:Pag-corrected =0.935,OR =0.98;rs60078183:Page-currected =0.782,OR =1.04;rs11204213:Page-currected =0.058,OR =1.66),and the M AF of rs1800813 was significantly higher in the high myopia group than that in the normal control group (Page-currected =0.001,OR =0.64).The genotype frequency of rs74225573,rs60078183 and rs11204213 was not evidently different in additive model 1 (AB vs.BB),additive model 2 (AA vs.BB),dominant model (AA+AB vs.BB) and recessive model (AA vs.AB+BB) (all at P＞0.05),while significant differences were found in genotype frequency of rs1800813 both in additive model 1 and dominant model (additive model 1:P=0.002,OR=0.59;dominant model:P=0.001,OR=0.58).Conclusions The SNP of rs1800813 in the PDGFRA gene is associated with the pathogenesis of high myopia in the Chinese Han population,but the SNPs of rs74225573 (MTOR gene),rs60078183 (CMPK1 gene) and rs11204213 (RBP3 gene) appear to be not associated with high myopia.

Objectives To investigate the prenatal diagnosis method of spinal muscular atrophy with amniotic fluid sample.Methods Totally 1 064 amniotic fluid samples from mid-trimester pregnant women were enrolled during January 2015 and January 2016 in 4 hospitals.Genetic analysis was performed for detecting potential contamination of maternal tissue by a genetic technique based on short tandem repeat ( STR) markers.Deletion of SMN1 gene was detected in 1 062 uncontaminated amniotic fluid samples by real-time PCR and multiplex ligation-dependent probe amplification ( MLPA) respectively.Results Two contaminated amniotic fluid samples were detected within 1 064 mid-trimester pregnant women by STR genotyping.The other 1 062 uncontaminated amniotic fluid samples were tested by real-time PCR.There were 37 samples with heterozygous deletion of Exon 7 of SMN1 gene ( 3.67%) , 34 samples with heterozygous deletion of Exon 8 of SMN1 gene (3.2%) and two samples with homozygous deletion of Exon 7 and Exon8 of SMN1 gene ( 0.19%) respectively , while other samples observed with no deletion of Exon 7 and Exon8 in SMN1 gene.Totally 41 samples with heterozygous or homozygous deletion of SMN 1 gene and 55 samples with undetected deletion of SMN 1 gene were confirmed by MLPA and the results showed 100%consistence with that of real-time PCR.Conclusions Both real-time PCR and MLPA are suitable for detecting the deletion of SMN 1 gene with amniotic fluid sample . Real-time PCR exhibits less sample requirement and time compared with MLPA .

Objective To provide data for reference on the impact of cyclosporin A (CsA) on the proliferation of the bone marrow mesenchymal stem cells in MDS patients through the investigation of the impact of cyclosporin A on human bone marrow mesenchymal stem cell proliferation. Methods The absorption rates of the bone marrow mesenchymal stem cells in the control group and the MDS patient group were determined by using the tetrazolinm salt (MTT) micro-colorimetric enzyme reaction. The concentrations of cyclosporine A are 2.5×10~4 ng/μl, 2.5×10~3 ng/μl, 2.5×10~2 ng/μl and 2.5×10ng/μl respectively. Results There is no significant difference between the each result by using different concentrations of CsA., which indicates the impact of CsA on the growth of mesenchymal stem cells is not significant(P >0.05). In the serial of concentrations mentioned, no cytotoxicity of CsA is observed. However, there is difference between the selected indicators of the control group and the patient group (P <0.01), and the value of the MDS patient group is higher than that of the control group. There is no statistic difference between the concentration of CsA and the data obtained from interactions between different groups (P >0.05). There is no significant difference between the absorption rates of the group treated by CsA of every concentration and the corresponding control group. Conclusion The impact of CsA on the bone marrow mesenchymal stem cell proliferation is significant in neither healthy people nor MDS patients.

Aim: To explore the in situ intestinal absorption in rats of ZLR-8, an insoluble NO-donor drug, and to compare the intestinal absorption enhancement by spray-dried emulsion. Methods: Intestine of rats was cannulat-ed for in situ perfusion. UV and HPLC methods were used to monitor phenolsulfonphthalein and ZLR-8, respec-tively. The effects on ZLR-8 absorption of the intestinal segments, the concentration of ZLR-8 and the pH of the circulating perfusate were studied. The absorption of ZLR-8 suspension was compared to that of the spray-dried emulsion. Results: 1-h in situ intestinal perfusion of the spray-dried emulsion allowed the estimation of the absor-tion percentage to be (23. 54 ± 1. 40) %, (15. 95 ± 0. 09) %, (12. 30 ± 0. 74) %, (3. 98 ± 0. 12) %, respec-tively; the absorption rate constants in duodenum, colon, jejunum and ileum to be (0.248 6 ±0.046 0) h~(-1), (0. 143 7 ±0. 036 0) h~(-1), (0. 069 2 ±0. 001 3) h~(-1), (0. 020 8 ±0. 000 4) h~(-1), respectively. Significant differ-ences in absorption characteristics were found among intestinal segments. In the range of 3. 4-9. 4, pH of the per-fuate had significant influence on the absorption of ZLR-8, and better absorption appeared at pH of 5. 4 to 7. 4. It was found that the absorption rate constant was unaffected by ZLR-8 concentration. However, the absorption amount was proportional to ZLR-8 concentration. Compared to the ZLR-8 suspension, the in situ intestinal absorption of ZLR-8 in rats given the spray-dried emulsion increased significantly. Conclusion: It was only found that ZLR-8 administered in suspension has minor absorption in rat duodenum while no apparent absorption occurred in other segemnts. ZLR-8 in spray-dried emulsion was fairly absorbed in the rat intestinal segments. Passive diffusion was invloved in the absorption of ZLR-8. Spray-dried emulsion significantly enhanced the intestinal absorption of ZLR-8 in rats.

0.05),but the level of P- selectin in treatment group were obviously decreased compared with control group after the seventh and fourteenth day(P

Objective To discuss the diagnosis and treatment of normotensive pheochromocyto-ma. Methods The clinical data of 22 patients with normotensive pheochromocytoma were reviewed. Inclusion criteria for normotensive pheochromocytoma were no previous history of hypertension and episode of symptoms suggesting high blood pressure. The blood pressure on admission was 90-130/ 60-90 mm Hg with an average of 113/72 mm Hg. Seven patients were found adrenal mass by routine ultrasonic examination. Twelve patients presented with superior abdominal or flank pain. Four pa-tients were present with fatigue, and 2 patients had fever. Headache and palpitation were found in 1 patient. Most of patients were present with large and round mass with low density area in the center of the tumor by uhrosonography and CT. Four patients had elevated level of plasma epinephrine and nor-epinephrine. 24 hours urine CA and VMA were elevated in 5 and 4 patients respectively. Seven pa-tients were prepared with infusion preoperatively to expand intravascular volume, and 2 patients were given prazosin 1.5 mg/d for 5 to 7 days. Results During the operation, seventeen patients had ele-vated blood pressure and 5 patients had no changed. One of seven patients with preoperative prepara-tion had obvious hypertension during operation, and 11 of 15 patients without preoperative preparation had obvious hypertension. The tumors were removed successfully in 21 patients. All the patients were diagnosed pheochromocytoma pathologically. Twenty-one patients had normal blood pressure with no recurrence during the follow-up from 1 month to 7 years. Conclusions The patients with normotensive pheochromocytomas may have lower catecholamine in their plasma and urine. The application of α-blockers and the expanding intravascular volume before operation could be important for the patients safe.

ObjectiveTo investigate the feasibility and efficacy of using the hepatoduodenal ligament tension-reduced operation (tension-reduced operation in short) for iatrogenic bile duct injury where the bile duct was severely defective. MethodsBetween March 2006 and May 2009, the authors treated 6 patients with iatrogenic bile duct injury (Bismuth type Ⅱ : 5 patients and type Ⅲ : 1 patient). A no. 7 black silk thread was used to hold the hilar plate tissues and the seromuscular layer of the bulbous part of the duodenum closer together and knots were tied. This method brought the porta hepatis and the duodenal bulb closer together and the hepatoduodenal ligament was shortened. An end to end anastomosis could then be made between the two broken ends of the defective bile duct without tension. ResultsSix patients suffered from bile duct injury and they recovered fully after the tensionreduced operation. There was no complication on follow-up. ConclusionsThe tension-reduced operation was efficacious in the treatment of iatrogenic bile duct injury. This technique should be popularized and more widely used.