Aim To investigate the effect of probucol on reverse cholesterol transport from macrophage to feces in vivo,serum lipid profiles of mice were tested before and after probucol administration for 4 weeks,and the 3H-contents in serum,liver and feces in mice were quantitated after 24 hours intraperitoneally injected macrophages which were labeled with 3H cholesterol.Methods 32 C57BL/6 mice were treated with different probucol(0,0.1%,0.5%,1% W/W)in chow diet for 4 weeks,then these mice were injected intraperitoneally with RAW264.7 macrophages(0.5 ml?mice,cells 5.0?106 with 6.2?106 cpm)which were loaded with cholesterol by incubation with acetylated LDL,labeled with 3H-cholesterol.Serum profiles were quantitated by enzymatic method;serum and liver tissues were harvested at 24th hour,feces were collected(0~24 hs),and all were analyzed for tracer counts(all were expressed as the percentage of the total injected counts per minute).Results After 4 weeks 0.1% probucol administration,the levels of total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C)decreased markedly by 34.3%,52.8%,(P

Objective Cardiac muscle cells play a critical role in maintaining normal function of the heart.Cardiotrophin-1(CT-1),a potent cardiac survival factor,is capable of inhibiting apoptosis or promoting survival in cardiomyocytes.To elucidate the mechanism of CT-1 promoting cardiac myocyte survival in cultured neonatal rat cardiomyocytes.To explore the potential signaling pathway that might be responsible for this effect.Methods We examined the cardiac myocyte survival effect of CT-1 in cultured neonatal rat cardiomyocytes.The cardiomyocytes were stained [3-(4,5-dimethyl-thiaziazol-2-yl)-2-5-diphenyltetrazolium bromide,MTT] and the counted.Results The survival rate of cardiac myocytes was increased by CT-1 in a dose-dependent manner(10-10～10-7 mol/L) and in a time-dependent manner(1～4 d,10-8 mol/L) in cultured neonatal rat cardiomyocytes.Pretreatment of PD098059(5?10-5mol/L),a MAPK blocker,decreased significantly survival rate of cardiac myocytes by promoted CT-1.The phorbol 12-myristate 13-acetate(PMA)(10-5mol/L),a PKC activator,increased significantly this effect of CT-1,but inhibited significantly by MAPK blocker PD098059.Conclusion CT-1 is a potent factor of promoting cardiac myocyte survival,and increase significantly survival rate of cardiac myocytes in a dose-dependentand a time-dependent manner in cultured neonatal rat cardiomyocytes.The MAPK signaling pathway mediates CT-1 induced cardiac myocyte survival.PKC signaling molecule may be a upstream signaling transduction pathway which cascades of MAPK in CT-1 induced cardiac myocyte survival.

Objective To investigate the clinical and biochemical metabolic features of 12 patients with systemic primary carnitine deficiency(CDSP) and to identify the SLC22A5 gene mutation types of the disease. Method The clinical and biochemical data were collected by retrospective analysis. DNA direct sequencing and multiplex ligation dependent probe amplification(MLPA)were applied for SLC22A5 gene analysis. Result Among 12 patients with CDSP, 3 cases had evident infection factors, 6 cases with convulsions, 5 cases manifested liver hypertrophy, 8 cases with hyperammonemia, and 9 cases showed myocardial damage. All CDSP patients were detected biallelic pathogenic mutation in SLC22A5 gene by direct sequencing. The gene types include IVS2+1G>T, c.3G>T(p.Met1Ile), c.760C>T(p.Arg254X), c.1400C>G(p.Ser467Cys), c.844dupc(p.Arg282fs), c.338G>A(p.Cys113Tyr), c.51C>G(p.Phe17Leu), c.659A>T(p.Glu220Val), and c.1365dupC(p.Thr456fs). c.659A>T(p.Glu220Val) and c.1365dupC(p.Thr456fs)are novel mutations. One female patient was maternal CDSP, her child had abnormal newborn screening. The allele frequency of c.760C>T(p.Arg254X) and c.1400C>G(p.Ser467Cys) were 37.5%(9/24)and 29.2%(7/24)respectively. The MLPA test results of all patients were negative. Conclusion The clinical manifestations are complex and various in patients with CDSP. Point and small InDel(insertions/deletions)mutation constitute the major alteration in SLC22A5 gene. c.1400C>G(p.Ser467Cys) might be another prevalence mutation type in Chinese CDSP patient.

OBJECTIVETo investigate the clinical phenotype and ACAT1 gene mutation in a family affected with beta-ketothiolase deficiency (BKTD).

METHODSClinical features and laboratory test data were collected. The probands were monozygotic twin brothers. Genomic DNA was isolated from peripheral blood leukocytes obtained from the probands and their family members. Molecular genetic testing of the ACAT1 gene was carried out.

RESULTSThe probands have presented with fever, vomiting and severe ketoacidosis. By arterial blood gas testing, pH was determined to be 7.164, bicarbonate was 4.0 mmol/L, and urine ketone was ++++. Urinary organic acid gas chromatography-mass spectrometry analysis showed excessive excretion of 3-hydroxybutyric acid, 2-methyl-3-hydroxybutyric acid and tiglylglycine. Increased 3-hydroxybutyrylcarnitine (C4-OH), tiglylcarnitine(C5:1) and 3-hydroxyisovalerylcarnitine (C5-OH) levels. The clinical phenotype of proband's parents were both normal, but an elder sister turned out to be an affected patient. Genetic analysis has identified two heterozygous mutations [c.622C>T(p.R208X) and c.653C>T (p.S218F)] in the proband, which were respectively detected in the mother and father. The c.653C>T (p.S218F) mutation was not found among the 100 healthy controls and has not been included in the Human Gene Mutation Database(HGMD).

CONCLUSIONThe primary clinical manifestations of BKTD is ketoacidosis. Urine organic acid and blood acylcarnitine analyses play an important role in the diagnosis of the disease. The compound heterozygous of ACAT1 gene mutations probably underlie the BKTD in our patient.

Acetyl-CoA C-Acetyltransferase ; genetics ; Acetyl-CoA C-Acyltransferase ; deficiency ; genetics ; Amino Acid Metabolism, Inborn Errors ; genetics ; Computational Biology ; Female ; Humans ; Infant ; Male ; Mutation ; Phenotype

OBJECTIVETo explore pathogenic mutation in a family affected with 2-hydroxyglutaric aciduria.

METHODSExons of 3 candidate genes, including L2HGDH, D2HGDH and SLC25A1, were amplified with polymerase chain reaction and subjected to direct sequencing.

RESULTSDNA sequencing has found that the proband and his affected younger brother have both carried a heterozygous mutation c.845G>A (p.R282Q) in the exon 7 of the L2HGDH gene. The same mutation was not detected in the his sister who was healthy. Pedigree analysis has confirmed that the above mutation was inherited from the mother. No mutation was detected in exons and flanking sequences of the D2HGDH and SLC25A1 genes.

CONCLUSIONMutation of the L2HGDH gene probably underlies the 2-hydroxyglutaric aciduria in this family.

Alcohol Oxidoreductases ; genetics ; Base Sequence ; Brain ; diagnostic imaging ; Brain Diseases, Metabolic, Inborn ; diagnostic imaging ; enzymology ; genetics ; Child ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Radiography ; Young Adult

OBJECTIVETo analyze the clinical features and mutation of MUT gene in a Chinese patient with isolated methylmalonic acidemia.

METHODSThe clinical characteristics and laboratory tests data were collected. Genomic DNA was extracted from peripheral blood leukocytes. The 13 exons and their flanking sequences of the MUT gene were amplified with polymerase chain reaction and subjected to direct DNA sequencing.

RESULTSThe patient has featured failure to thrive, lethargy, seizure, hypotonia, severe ketoacidosis and hyperammonemia. Tandem mass results showed reduction of multiple acylcarnitine. Urine organic acid testing showed pronounced increase in methylmalonate excretion. Homocysteine was normal. The patient showed no response to vitamin B12 treatment. The above results suggested that the patient had isolated methylmalonic acidemia. DNA sequencing analysis confirmed that the patient has carried two MUT gene mutations, c.755dupA and a novel mutation c.944dupT.

CONCLUSIONInherited metabolic disease screening plays an important role in the diagnosis of clinical diseases. However, to confirm the results will need gene mutation analysis.

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

OBJECTIVETo analyze PCCA and PCCB gene mutations in 10 Chinese patients with propionic acidemia(PA).

METHODSGenomic DNA was extracted from peripheral blood leukocytes. The 39 exons and flanking sequences of the PCCA and PCCB genes were amplified with polymerase chain reaction and subjected to direct DNA sequencing.

RESULTSDNA sequencing has revealed that 7 patients have carried a PCCA gene mutation, 2 patients carried PCCB gene mutation and 1 patient carried mutations in both PCCA and PCCB genes. Ten PA mutations were confirmed, including 8 affecting the PCCA gene and 2 affecting the PCCB gene. Three PCCA mutations c.245G>A, IVS15+5del5, c.1288C>T and 2 PCCB mutations c.838insC, c.1087T>C were found for the first time.

CONCLUSIONAmong Chinese patients with propionic acidemia patients, their genetic mutations are mainly found on the PCCA gene.

Child, Preschool ; Female ; Humans ; Infant ; Male ; Methylmalonyl-CoA Decarboxylase ; genetics ; Mutation ; Propionic Acidemia ; genetics

OBJECTIVETo identify the types of OTC gene mutations in three male patients with late onset ornithine transcarbamylase deficiency (OTCD, MIM #311250).

METHODSGenomic DNA was extracted from peripheral blood leukocytes. The 10 exons and their flanking sequences of the OTC gene were amplified with polymerase chain reaction and subjected to direct DNA sequencing.

RESULTSBased on DNA sequence analysis, all of the three patients have carried OTC gene mutations. Patients 1 and 2 were both hemizygous for mutation c.586G> A(p.D196N). A novel mutation c.800G> C(p.S267T) were confirmed in patient 3.

CONCLUSIONp.S267T mutation has affected the conserved amino acid motif of the OTC protein, and is therefore a pathogenic mutation.

Age of Onset ; Amino Acid Sequence ; Base Sequence ; Child ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Ornithine Carbamoyltransferase ; genetics ; Ornithine Carbamoyltransferase Deficiency Disease ; epidemiology ; genetics ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid

Objective:To investigate the clinical application effect of bypass vein bridging in repairing high-voltage electric burn wounds on the head with free anterolateral thigh flaps.Methods:This study was a retrospective observational study. From May 2017 to December 2022, 8 patients with high-voltage electric burns on the head who met the inclusion criteria were admitted to Zhengzhou First People's Hospital, including 6 males and 2 females, aged 33 to 73 years. All patients had skull exposure, including 3 cases of large skull defect, 1 case of left eye necrosis, and 3 cases of cerebral hemorrhage. After debridement, the head wound area was from 13 cm×7 cm to 21 cm×15 cm, and the free anterolateral thigh flap with the area of 14 cm×8 cm to 22 cm×16 cm was cut for repair. The main descending branch of the lateral circumflex femoral artery carried by the flap was anastomosed end-to-end with the superficial temporal artery in the recipient area. One accompanying vein of the anastomotic artery of the flap was end-to-end anastomosed with the branch of the external jugular vein via great saphenous vein bridging, and the other accompanying vein was end-to-end anastomosed with the superficial temporal vein in the recipient area. The donor site wounds were directly sutured or closed with medium-thickness skin grafts from inner thigh. The blood supply and survival of the flap, and the wound healing on the head were observed after operation. The blood flow and lumen filling of the transplanted vein were observed and recorded by using color ultrasound diagnostic system within 2 weeks after operation. The wound repair method and wound healing of the flap donor site were recorded and observed. Patients were followed up to observe the appearance of the flaps and the flap donor sites, the muscle strength of the lower limbs where the flap donor site was located, and whether the patient could complete standing, walking, and squatting using the lower limbs where the flap donor site was located.Results:The flaps of 8 patients survived after operation, and no arterial or venous crisis occurred. The wounds of 5 patients on the head healed after operation, and the wounds of 3 patients on the head healed after second debridement 21 to 35 days after operation due to exudates under the flap 2 weeks after operation. Within 2 weeks after operation, the grafted vein continued to be unobstructed. After the ultrasound probe was pressurized, the grafted vein could be deflated, and the blood vessels were rapidly filled after the probe was released. The wounds of flap donor sites of 3 patients were directly sutured and healed 2 weeks after operation. The wounds of flap donor sites of 5 patients were closed with medium-thickness skin grafts from inner thigh, and all the skin grafts survived 12 days after operation. During follow-up of 6 to 12 months, the head flaps of all patients were slightly bloated without hair growth. Mild linear or patchy scar hyperplasia was left in the donor site. The muscle strength of the lower limbs where the flap donor site was located was normal and did not decrease. The patients could stand, walk, and squat with the lower limbs where the flap donor site was located.Conclusions:When using the free anterolateral thigh flap to repair high-voltage electric burn wounds of various areas and depths on the head, bypass vein bridging can reduce the occurrence of postoperative flap vein crisis and improve the quality of postoperative wound healing without affecting the function of the lower limbs where the flap donor site is located.

OBJECTIVETo detect potential mutations of Y9ASS1, ASL and SLC25A13 genes in four patients manifesting citrullinemia.

METHODSGenomic DNA was extracted from peripheral blood leukocytes. Exons and their flanking sequences of the three genes were amplified with polymerase chain reaction and subjected to direct DNA sequencing.

RESULTSBased on DNA sequence analysis, one case was diagnosed with argininosuccinate synthetase deficiency, and the mutation type (ASS1 gene) was c.236C>T (p.S79F) + c.431C>G (p.P144R). Two cases were diagnosed with argininosuccinic aciduria (ASL gene), and their gene mutations were c.434A>G (p.D145G) + c.1366C>T (p.R456W) and c.331C>T (p.R111W) + IVS8+2insT, respectively. A thirteen months boy who carried a heterozygous 851del4 mutation (SLC25A13 gene) was diagnosed with citrullinemia adult-onset type II.

CONCLUSIONThrough analysis of relevant pathogenic genes, four patients have been diagnosed.

Adult ; Amino Acid Sequence ; Argininosuccinate Lyase ; genetics ; metabolism ; Argininosuccinate Synthase ; genetics ; metabolism ; Base Sequence ; Citrullinemia ; enzymology ; genetics ; Female ; Humans ; Infant ; Male ; Mitochondrial Membrane Transport Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Pedigree ; Point Mutation