Objective To screen out the tetrahydrobiopterin def iciency in patients with hyperphenylalaninemia by urinary pterin analysis. Methods Analysis of urinary neopterin (N) and biopterin (B) was done in 96 patients with hyperphenylalaninemia by high performance liquid chromatography. Combined BH4 loading test was performed for patients with abnormal urinary pterin profiles. Results Eleven patients were diagnosed as ha ving BH4 deficiency caused by 6 pyruvoyl tetrahydrobiopterin synthase deficiency. They had a much high N/B ratio(N/B:197?250) and a very low B percentage (1.0?0.8)%. Blood phenylalanine levels were decreased from (720 1 200) ?mol/L to (120 240) ?mol/L after taking BH4 tablets in 4 of 11 patients. Conclusion The incidence of BH4 deficiency among hyperphenylalaninemia is about 12%. Analysis of urinary pterin is feasible and effective in screening of BH4 deficiency.

Objective Maple syrup urine disease (MSUD) is a rare metabolic disorder caused by deficiency of the activity of branched-chain 2-keto acid dehydrogenase complex.The complex contains E1α,E1β and E2 subunits which are encoded by BCKDHA,BCKDHB or DBT genes respectively.Mutation in any gene will cause MSUD.The aim of this study was to analyze the gene mutations of four cases with MSUD and carry out prenatal diagnosis for these four families for MSUD.Methods From 2005 to 2010,four neonates (two males and two females) were diagnosed as MSUD at 2,5,10and 26 days of life.The coding regions of BCKDHA gene and BCKDHB gene in the above four cases were amplified by polymerase chain reaction and analyzed by direct DNA sequencing.During the second pregnancy of the same mother,the amniotic fluid was drawn out at 16-20 weeks for gene mutation analysis after the amniocytes were cultured.Results Mutation analysis revealed six mutations in four patients,including four novel mutations (c.308T＞C,c.562G＞T,c.1279C＞G and c.1280-1291de112) and two previously reported mutations.Five mutations (c.308T＞C,c.562G ＞T,c.868G＞A,c.1279C＞G and c.1280-1291de112) were detected on BCKDHA gene in three patients.While one mutation (c.853C＞T) was found on BCKDHB gene in one patient.Only one mutation was found in the amniocytes of each patient's mother at their second pregnancies suggesting a MSUD heterozygous fetus.Conclusions Analysis of BCKDHA and BCKDHB allowed preliminary understand of gene mutations in the four MSUD families,and made prenatal diagnosis possible,which helped in consultation in the second pregnancy.

ld have an improved outcome after reasonable treatments. The gene mutation detection suggests that 609G>A (W203X) may be the hot spot mutation of MMACHC gene in Chinese patients.

OBJECTIVESTo assess the incidence of tetrahydrobiopterin (BH4) deficiency among patients with hyperphenylalaninemia (HPA) in southern Chinese and evaluate clinical outcome and gene mutations in tetrahydrobiopterin deficient patients.

METHODSUrinary neopterin (N) and biopterin (B) was analyzed in 87 patients with hyperphenylalaninemia by high-performance liquid chromatography. Further combined loading tests with phenylalanine (Phe) (100 mg/kg) and tetrahydrobiopterin (BH4) (7.5 mg/kg) were performed in suspected patients with abnormal urinary pterin profiles. Gene mutation analysis was performed for patients with BH4 deficiency and their parents. BH4 deficient patients were treated with BH4 and neurotransmitter precursors after diagnosis. Blood phenylalanine levels, clinical symptoms and mental development were followed up.

RESULTSEleven patients were diagnosed as having BH4 deficiency caused by 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency. The incidence of tetrahydrobiopterin (BH4) deficiency among patients with hyperphenylalaninemia (HPA) in southern Chinese was 10%. Combined loading tests with phenylalanine and oral BH4 were done in 4 of 11 patients and their phenylalanine levels were decreased to normal 4 - 6h after BH4 administration. Four different mutations (P87S, N52S, D96N and G144R) in the PTPS gene were detected in 5 families. Five PTPS-deficient patients were treated with synthetic BH4, neurotransmitter precursors (L-dopa plus carbidopa, and 5-hydroxytryptophan). They had satisfactory physical and mental development after treatment. One patient with partial PTPS deficiency had normal growth and mental development without treatment.

CONCLUSIONSOur results emphasize that screening for BH4 deficiency should be carried out in all patients with hyperphenylalaninemia in order to minimize the misdiagnosis. Patients with BH4 deficiency should be treated early with BH4 and a combination of neurotransmitter precursors.

Biopterin ; administration & dosage ; analogs & derivatives ; deficiency ; urine ; China ; DNA Mutational Analysis ; DNA, Complementary ; chemistry ; genetics ; Follow-Up Studies ; Genetic Testing ; Humans ; Mutation, Missense ; Neopterin ; urine ; Phenylketonurias ; blood ; enzymology ; genetics ; Phosphorus-Oxygen Lyases ; genetics ; metabolism

OBJECTIVETo investigate the MUT gene mutations in patients with methylmalonic acidemia (MMA), and analyze the genotype-phenotype correlation in patients with methylmalonyl-CoA mutase deficiency.

METHODSThe diagnosis of the disease mainly depends on the measurement of C3 (acylcarnitine), C3/C0 (free carnitine) and C3/C2 (acetylcarnitine) in the blood by tandem mass spectrometry, the detection of methylmalonic acid in the urine by gas-chromatography mass spectrometry, the determination of total homocysteine in the serum, and the loading test of vitamin B(12). The entire coding region of the MUT gene was screened by PCR combined with direct DNA sequencing in 21 isolated MMA patients. Novel mutations were identified by restriction fragment length polymorphism (RFLP) and sequence analysis in 100 controls.

RESULTSSeventeen MUT gene mutations were detected in 14 of the 21 patients, among them 8 mutations were novel, and R108H, D244LfsX39 and G544X were more frequent, with the frequencies of 9.5%, 7.1% and 9.5%, respectively. Most mutations were missense mutations (64.7%), and majority of them were in exons 2 and 3 (55.6%). Ten out of the 14 patients with MUT gene mutations had early-onset disease, while one case had late-onset disease, and the remaining 3 cases were detected by newborn screening. In addition, 11 of these 14 patients did not respond to vitamin B(12).

CONCLUSIONThis study revealed partial MUT gene mutation spectrum in Chinese patients with isolated MMA. The patients carrying MUT mutations often had early-onset disease, and most of them were VitB(12)- non-responsive.

Amino Acid Metabolism, Inborn Errors ; genetics ; Base Sequence ; China ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Methylmalonic Acid ; metabolism ; Methylmalonyl-CoA Mutase ; genetics ; Molecular Sequence Data ; Mutation

Objective To compare the safety of octogenarian patients receiving non-cardiac surgery within 1 week versus within 1-3 weeks after coronary computed tomographic angiography(CTA).Methods Octogenarian patients who underwent non-cardiac surgery after coronary CTA in Fujian Medical University Union Hospital,were retrospectively analyzed.All patients were divided into two groups:those received surgery within 1 week after coronary CTA as group 1 (n =73),those within 1-3 week after coronary CTA as group 2,(n =35).The baseline clinical characteristics,the changes in pre-and postoperative serum creatinine levels (Scr)and estimated glomerular filtration rate(eGFR),and the incidence of acute kidney injury(AKI)were compared between two groups.The revised cardiac risk index(RCRI)score was evaluated for each octogenarian inpatient,and the RCRI sum score for different types of non-cardiac surgery were calculated.Finally,the RCRI sum score of the preoperative risk factors were compared between different types of non-cardiac surgery so as to assess their specifically safety.Results In 108 patients who performed coronary CTA,only one patients developed palpitation and three had injection site pain.All patients receiving different types of non-cardiac operation had low revised cardiac risk index(RCRI ≤ 2).Death was not found.The serum levels of Scr and eGFR were similar between two groups before coronary CTA and after operation(all P＞0.05).Conclusions Octogenarian patients with low preoperative cardiac risk index(RCRI ≤ 2) are safe for performing non-cardiac surgery within 1 week after coronary CTA.

Objective:To investigate the effect of autophagy related gene Atg101 on white adipocyte senescence.Methods:An Atg101 knockdown model of 3T3-L1 mature adipocytes was constructed to probe the effect of Atg101 on autophagy-related proteins LC3 and p62 protein. The RNA-seq database of human subcutaneous adipose tissue was constructed and analyzed, and the co-expressed gene set was predicted based on the pearson correlation coefficient( R2>0.4, P<0.05) between FPKM values of Atg101 and other gene, followed by KEGG and Reactome enrichment analysis. Young mouse(8 weeks old) and old mouse(18 months old) models were established, and the expression levels of Atg101 in inguinal white adipose tissue and epididymal white adipose tissue were detected by quantitative real-time PCR(RT-qPCR) and Western blot. Furthermore, the differences in white adipocyte senescence-associated secretory phenotype(SASP), cell cycle and mitochondrial homeostasis-related genes were detected by RNA-seq, Western blot, and RT-qPCR to analyze the effects of Atg101 silencing on adipocyte senescence. Results:The autophagy-related protein LC3-Ⅱ expression was significantly decreased and p62 protein was induced after Atg101 was knockdowned in 3T3-L1 adipocytes, suggesting impaired cell autophagy. KEGG enrichment analysis revealed that Atg101 co-expressed gene set was mainly enriched in autophagy and senescence-related pathways; Reactome enrichment analysis revealed that this gene set was associated with multiple cell cycle signaling pathways. RT-qPCR and Western blot confirmed that both mRNA and protein levels of Atg101 were down-regulated in inguinal white adipose tissue of aging mice, and protein levels in epididymal white adipose tissue were also significantly reduced. Finally, it was further confirmed that SASP-related genes were induced after Atg101 knockdown in white adipocytes, and cell cycle-specific gene expression was restricted and cytokine-dependent protein kinase inhibitors p16 and p21 expressions were significantly increased, while mitochondrial homeostasis regulatory genes were also suppressed.Conclusions:Knockdown of Atg101 may regulate white adipocyte senescence by inhibiting autophagic activity, presenting impaired mitochondrial homeostasis.

Objective To explore the therapeutic effects of estrogen-intervened endothelial progenitor cells( EPCs) transplantation on diabetic ischemic stroke rats. Methods PKH26-labeled diabetic EPCs and estrogen-intervened diabetic EPCs were injected into rats via the tail vein 24 h after cerebral ischemia. Cerebral ischemic volume, behavioral changes, ischemic site vascularization and homing of EPCs were measured 3 d after EPCs injection. Results Compared with diabetic ischemic rats, estrogen-intervened EPCs transplantation had reduced infarct volumes, improved behavioral scores and ischemic site revascularization and promoted homing of EPCs to sites of injury(P<0.05). Conclusion Estrogen-intervened EPCs transplantation had a better therapeutic effect on diabetic ischemic stroke by promoting EPCs homing to injury site and EPCs-medicated neovascularization .

OBJECTIVETo analyze the clinical characteristics of the patient with tyrosine hydroxylase deficiency, and investigate it's molecular mechanism.

METHODThe clinical characteristics of a patient with tyrosine hydroxylase deficiency were summarized and analyzed, his and his family's peripheral blood specimens were collected after informed consent was signed. All exons and the intron-exon boundaries of guanosine triphosphate hydroxylase I gene, tyrosine hydroxylase gene and sepiapterin reductase gene were examined by DNA-PCR, bi-directional sequencing.

RESULTThe patient was a 3-year-old boy, presented with unexplained dystonia for 3 years, without significant impairment of intelligence. Physical examination showed limb muscle strength grade V, rigidity of extremities, hypertonicity, brisk deep tendon reflexes in limbs, without obvious abnormalities in auxiliary examination, such as brain MRI, hepatic biochemical panel, creatine kinase, and ceruloplasmin. He dramatically responded to small doses of levodopa in the follow-up for half a year. A homozygous missense change in exon 5 of TH gene, c.605G > A (p.R202H), which was a known pathogenic mutation, was found in the patient. His parents were heterozygous for the R202H mutation.

CONCLUSIONThe age of onset in tyrosine hydroxylase deficiency patients is usually within the first year of life. Unexplained dystonia and hypokinesia were the main clinical features of tyrosine hydroxylase deficiency. The dopa-responsive effects for some patients are so obvious that we should strengthen awareness of the disease. TH gene c.605G > A (p.R202H) may be a common type of causative mutations for the mild form at home and abroad.

Brain ; metabolism ; pathology ; Catecholamines ; biosynthesis ; Child, Preschool ; DNA ; genetics ; DNA Mutational Analysis ; Dopamine Agents ; administration & dosage ; therapeutic use ; Dystonic Disorders ; drug therapy ; genetics ; metabolism ; Homozygote ; Humans ; Hypokinesia ; drug therapy ; genetics ; metabolism ; Levodopa ; administration & dosage ; therapeutic use ; Male ; Muscle Rigidity ; drug therapy ; genetics ; metabolism ; Mutation, Missense ; Polymerase Chain Reaction ; Tyrosine 3-Monooxygenase ; deficiency ; genetics ; metabolism