Clinical application study on capillary electrophoresis-based gene diagnosis of hyperphenylalaninemia / 中华实用儿科临床杂志

Objective:To evaluate the clinical value of a capillary electrophoresis-based method for gene diagnosis of hyperphenylalaninemia.Methods:In this single-center prospective study, 40 newborns with suspected hyperphenylalaninemia detected by neonatal liquid chromatography-tandem mass spectrometry screening at Nanjing Maternity and Child Health Care Hospital from February 2021 to February 2023 were included, with 22 males, 18 females and a mean age at diagnosis of 21.93 days.Capillary electrophoresis was used to detect 85 variants of the phenylalanine hydroxylase ( PAH) gene in 40 newborns with suspected hyperphenylalaninemia.The PAH gene of undiagnosed patients was further analyzed by Sanger sequencing.The detection rate, sensitivity and specificity of capillary electrophoresis were calculated. Results:Among these 40 newborns with suspected hyperphenylalaninemia, 71 PAH variants were detected by capillary electrophoresis, 32 patients were clearly diagnosed, only 1 pathogenic variant was found in 5 patients, and no pathogenic variant was found in the last 3 patients.Therefore, the detection rate, sensitivity and specificity of capillary electrophoresis for analysis of the PAH gene were 80.00%, 88.75% and 100%, respectively. Conclusions:The capillary electrophoresis-based method can rapidly, efficiently and accurately detect PAH gene variants at lower cost and is a promising gene detection method for hyperphenylalaninemia in clinical practice.

Effect of PACS-2 on the development of Alzheimer's disease / 中国现代医生

@#Objective To investigate the involvement of phosphofurin acidic cluster sorting protein-2(PACS-2)in mitochondrial function and apoptosis in N2a/APP695swe cells and further explore the role and significance of PACS-2 in the development of Alzheimer's disease(AD).Methods The CCK8 method was used to analyze the cell survival rate of N2a/APP695swe cells treated with different concentrations of tetrahydroxy stilbene glycoside(TSG)for 48h and to select the appropriate concentration of TSG for subsequent experiments.N2a/WT cells and N2a/APP695swe cells were routinely cultured in vitro,and the experimental cells were divided into 3 groups:blank control group(WT group):N2a/WT cells;model group(APP group):N2a/APP695swe cells;treatment group(TSG group):N2a/APP695swe cells with appropriate concentrations of TSG intervention.TUNEL method to observe apoptosis by fluorescence microscopy;JC-1 method for flow detection of cellular mitochondrial membrane potential;WB to detect protein expression of PACS-2;RT-qPCR to detect PACS-2 mRNA expression.Results CCK8 method was used to analyze the cell survival rate of different concentrations of TSG acting on cells after 48h:the protective effect of 100 μmol/L TSG was the most significant and the difference was statistically significant(P<0.01).The TUNEL method of fluorescence microscopy observed the apoptosis:compared with the WT group,the apoptosis rate of APP group was increased,compared with the APP group,the apoptosis rate of TSG group was decreased,and the differences were statistically significant(P<0.05).The JC-1 method was used to detect the mitochondrial membrane potential of cells:compared with the WT group,the membrane potential of APP group was decreased,compared with the APP group,the membrane potential of TSG group was increased,and the differences were statistically significant(P<0.05);Western blot(WB)detection of PACS-2 protein expression:compared with the WT group,PACS-2 expression was significantly higher in the APP group,and compared with the APP group,PACS-2 expression was significantly lower in the TSG group,with statistically significant differences(P<0.05);The RT-qPCR detected the mRNA expression of PACS-2:the expression of PACS-2 was elevated in the APP group compared with the WT group and decreased in the TSG group compared with the APP group,with statistically significant differences(P<0.05).Conclusion PACS-2 has an important role in the development of AD,and its upregulation may promote the development of AD.The cerebroprotective drug TSG may exert cytoprotective effects by downregulating PACS-2 to inhibit apoptosis and improve mitochondrial function in AD model cells.

Application value of C14∶1/C12∶1 for very long chain acyl-CoA dehydrogenase deficiency in newborn screening / 中华检验医学杂志

Objective:To establish the cut-off value of tetradecenoyl carnitine (C14∶1)/dodecenoyl carnitine(C12∶1) based on non-derivatized tandem mass spectrometry (MS/MS), and to explore the application value of C14∶1/C12∶1 to screen newborns for very long chain acyl-CoA dehydrogenase deficiency (VLCADD), determining the best combination of indicators for screening VLCADD.Methods:This retrospective study included data from 17 newborns with VLCADD detected by MS/MS and confirmed by acyl-CoA dehydrogenase very long chain ( ACADVL) gene detection, and 423 507 newborns with normal MS/MS results. The data from these newborns were collected from January 2014 to December 2021 as the newborns received neonatal screening in Nanjing Neonatal Disease Screening Center and Suzhou Neonatal Disease Screening Center. All newborns were divided into 3 groups: all newborns group, full-term newborns group and normal-birth-weight newborns group, and the cut-off values of C14∶1/C12∶1 for VLCADD in these 3 groups were determined by their receiver operating characteristic (ROC) curves individually. With these results, a total of 5 interpretation schemes were composed using different indicators alone or jointly: scheme 1 being C14∶1/C12∶1, scheme 2 being C14∶1, scheme 3 being C14∶1+C14∶1/C2+C14∶1/C16, scheme 4 being C14∶1/C12∶1+C14∶1, and scheme 5 being C14∶1/C12∶1+C14∶1+C14∶1/C2+C14∶1/C16. The detection rate, false-positive rate and positive predictive value of each scheme were calculated, and their screening efficiencies were statistically compared by Chi-square tests. Results:The cut-off values of C14∶1/C12∶1 for VLCADD in the 3 newborn groups were all 2.80. The detection rates of VLCADD with all 5 interpretation schemes were 17/17. Scheme 1 had the highest false positive rate [26.15‰ (11 075/423 524)] and the lowest positive predictive value [0.15% (17/11 092)]. Scheme 4 (Scheme 5) had the lowest false positive rate [0.02‰ (10/423 524)] and the highest positive predictive value [62.96% (17/27)]. Comparing scheme 4 (Scheme 5) with scheme 1, scheme 2 and scheme 3, the differences of false positive rate (χ2=302.30,11 191.50,32.06) and positive predictive value (χ2=102.51,3 485.61,13.83) were statistically significant (all P<0.001). Conclusion:C14∶1/C12∶1 was an effective auxiliary interpretive indicator for VLCADD in newborn screening, and the combination of C14∶1/C12∶1+C14∶1 was tested to be the best indicator for VLCADD screening based on non-derivatized tandem mass spectrometry.

Effect of Jiaomu Gualou Decoction combined with bisoprolol on myocardial microcirculation and oxidation/antioxidant balance in patients with heart failure / 国际中医中药杂志

Objective:To investigate the effect of Jiaomu Gualou Decoction combined with bisoprolol on myocardial microcirculation and oxidative/antioxidant balance in patients with heart failure.Methods:A total of 128 patients with heart failure who met the inclusion criteria from March 2020 to February 2021 in Dagang Hospital of Traditional Chinese Medicine, Binhai New Area, Tianjin were divided into 2 groups by random number table method, with 64 in each group. On the basis of conventional treatment, the control group was given oral bisoprolol, and the study group was given Jiaomu Gualou Decoction and oral bisoprolol. Both groups were treated continuously for 4 weeks. TCM syndrome scores were performed before and after treatment. The left ventricular ejection fraction (LVEF), left ventricular end-systolic diameter (LVESD), and left ventricular end-diastolic diameter (LVEDD) were detected by ultrasonic diagnostic equipment, and the frequency, duration and total myocardial ischemia load of the 24-hour ECG were recorded. Lipid peroxide (LPO) was detected by fluorescence method, SOD, MDA and GSH-Px were detected by colorimetric method. The adverse events were recorded and clinical response was evaluated.Results:The response rate was 93.75% (60/64) in the study group and 79.69% (51/64) in the control group, and the difference between the two groups was statistically significant ( χ2=5.49, P=0.019). After treatment, the scores of shortness of breath, phlegm, sternocostal fullness, and fatigue in the study group were significantly lower than those in the control group ( t values were 8.48, 8.15, 8.86, and 6.88, respectively, all Ps<0.001). After treatment, the LVEF of the study group [(53.26±5.18)% vs. (48.65±5.27)%, t=4.99] was significantly higher than that of the control group, and the LVESD [(42.59±3.26) mm vs. (46.98±3.55) mm, t=7.29], LVEDD [(52.79±4.15) mm vs. (57.48±4.60) mm, t=6.06] significantly lower than the control group ( P<0.01). After treatment, frequency of ST segment fall [(2.51±0.42) times/24 h vs. (3.79±0.55) times/24 h, t=14.80], duration [(15.26±3.45) min/24 h vs. (22.65±3.71) min/24 h, t=11.67] and total myocardial ischemia load [(25.79±5.13) mm/min vs. (38.02±5.44) mm/min, t=13.09] were significantly lower than those in the control group ( P<0.01). After treatment, the levels of serum GSH-Px and SOD in the study group were significantly higher than those in the control group ( t values were 10.97, 14.37, respectively, all Ps<0.001), while the levels of LPO and MDA were significantly lower than those in the control group ( t values were 7.50, 11.04, respectively, all Ps<0.001). During the treatment period, the incidence of adverse events was 4.69% (3/64) in the study group and 7.81% (5/64) in the control group, with no significant difference between the two groups ( χ2=0.13, P=0.715). Conclusion:The Jiaomu Gualou Decoction combined with bisoprolol can improve the cardiac function and myocardial microcirculation in patients with heart failure, promote the body's oxidation/antioxidant balance, relieve the clinical symptoms of patients, and improve the response effect safely.

Tandem mass spectrometry and genetic variant analysis of four neonates with very long chain acyl-coenzyme A dehydrogenase deficiency / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical features and genetic variants in four neonates with very long chain acyl-coenzyme A dehydrogenase (VLCAD) deficiency.@*METHODS@#Neonates with a tetradecenoylcarnitine (C14:1) concentration at above 0.4 μmol/L in newborn screening were recalled for re-testing. Four neonates were diagnosed with VLCAD deficiency by MS-MS and genetic testing, and their clinical features and genotypes were analyzed.@*RESULTS@#All cases had elevated blood C14:1, and the values of first recalls were all lower than the initial test. In 2 cases, the C14:1 had dropped to the normal range. 1 case has remained at above 1 μmol/L after the reduction, and the remainder one case was slightly decreased. In total eight variants of the ADACVL genes were detected among the four neonates, which included 5 missense variants and 3 novel variants (p.Met344Val, p.Ala416Val, c.1077+6T>A). No neonate showed salient clinical manifestations.@*CONCLUSION@#Above findings have enriched the spectrum of ADACVL gene mutations and provided a valuable reference for the screening and diagnosis of VLCAD deficiency.

Phenotypes and pathogenic variations in two cases of propionic acidemia / 中华围产医学杂志

Objective:To investigate the clinical characteristics and pathogenic mutations of propionic acidemia.Methods:Clinical data of two patients with propionic acidemia admitted to the Obstetrics and Gynecology Hospital of Nanjing Medical University from May 2017 to June 2018 were collected. Genomic DNA was extracted from the peripheral blood of the patients and their parents. Inherited disease panel based on Ion Torrent semiconductor sequencing technology was performed to detect gene mutations, and those with suspected pathogenic mutations were verified by Sanger sequencing. Descriptive statistical analysis was used for data analysis.Results:Case 1 was suspected of sepsis and admitted to the Obstetrics and Gynecology Hospital of Nanjing Medical University due to "drowsiness and milk rejection" on the second day after birth. Tandem mass spectrometry suggested the level of propionyl carnitine and its ratios to acetylcarnitine and free carnitine were increased. Urine gas chromatography-mass spectrometry showed elevated 3-hydroxypropionic acid and methylcitric acid. Genetic analysis revealed that the infant carried c.331C>T (p.R111X)/c.1228C>T (p.R410W) compound heterozygous mutations in the PCCB gene. The infant was diagnosed with propionic acidemia and treated with a special diet with an L-Carnitine supplement but died of sudden coma and vomiting without precipitating factors at three months of age. Case 2 presented with sudden vomiting, drowsiness, and anergia on the admission at five-months old. Tandem mass spectrometry showed increased propionyl carnitine level and its ratios. Compound heterozygous mutations of c.146delG (p.G49EfsX16)/c.1253C>T (p.A418V) in the PCCB gene were identified in the patient, of which c.146delG (p.G49EfsX16) was a de novo mutation and was evaluated as a pathogenic mutation. The patient was on a special diet with an L-Carnitine supplement, but with disobedience. Followed up to the age of three years and eight months, the child was severely underdeveloped. Conclusions:Neonates with clinically suspected sepsis may have propionic acidemia, and tandem mass spectrometry and genetic testing should be performed as soon as possible to confirm or rule out the diagnosis. Further investigations on the pathogenesis and function of the new mutation are still needed.

Prenatal management of multifetal pregnancies containing monochorionic fetuses / 中华围产医学杂志

Multifetal pregnancies with monochorionicity are more complicated, for which pregnancy monitoring and intrauterine intervention are of great importance. For dichorionic triamniotic triplets with MC twin, multifetal pregnancy reduction measures included cardiac injection of potassium chloride at 11 to 14 weeks of gestation for reduction to monochorionic singleton, and radiofrequency ablation after 16 weeks of gestation to preserve the dichorionic diamniotic twins. Moreover no significant difference was observed in the pregnancy outcomes between the two methods. Fetalscopic laser surgery can significantly improve the perinatal prognosis of multiple pregnancies complicated by twin-to-twin transfusion syndrome. Umbilical occlusion and transection can be used for the reduction of triplets containing monochorionic monoamniotic twins to avoid fetal death caused by entanglement of the umbilical cord. Cardiac injection of potassium chloride is appropriate for reducing the two fetuses in the same chorionic sac for trichorionic quadruamniotic pregnancy. Selective fetal reduction is applicable for MC triplets or quadruplets pregnancy, however, relevant studies are all with a small sample size, which requires full consultation and individualized treatment.

Preliminary screening of therapeutic drugs for phenylalanine hydroxylase deficiency / 中国基层医药

Objective:To screen new drugs for treatment of phenylalanine hydroxylase deficiency.Methods:From October 2019 to October 2020, virtual drug screening was performed in Center of Genetic Medicine, Nanjing Maternity and Child Health Care Hospital, Women's Hospital of Nanjing Medical University computer according to the characteristics of the binding ability of phenylalanine hydroxylase to drug spatial structure. Ten candidate drugs were screened from the FDA drug library (including 2 697 kinds of active pharmaceutical ingredients). A eukaryotic expression system was used to determine the effects of drugs on the activity of phenylalanine hydroxylase at the molecular level. Drug-sensitive mutants were screened.Results:Among the 10 candidate drugs, neoplasm hydrochloride, fluocinonide acetate and risperidone increased 23% [ t = 18.21, P < 0.001, vs. non-drug-treated phenylalanine hydroxylase group (i.e., only solvent and no drug added to the reaction system)], 21% ( t = 3.44, P < 0.05, vs. non-drug-treated phenylalanine hydroxylase group), 31% ( t = 19.57, P < 0.001, vs. non-drug-treated phenylalanine hydroxylase group) of the activity of phenylalanine hydroxylase. The remaining drugs exhibited weak even inhibitory effects on the activity of phenylalanine hydroxylase. 25% of p.D101N mutant could be activated by risperidone ( t = 15.86, P < 0.001, vs. non-drug-treated p.D101N mutant group). Conclusion:Neoplasm hydrochloride, fluocinonide acetate and risperidone can be used as potential therapeutic drugs for phenylalanine hydroxylase deficiency, and p.D101N mutant can be used as the drug-sensitive mutation site.

Etiological diagnosis and clinical evaluation of isolated fetal ascites / 中华妇产科杂志

Objective:To explore the correlation between prenatal clinical data with etiological diagnosis and neonatal outcome in isolated fetal ascites.Methods:Totally, 36 pregnancy cases diagnosed as isolated fetal ascites by ultrasound in Provincial Hospital Affiliated to Shandong University from June 22nd, 2016 to September 28th, 2018 were collected. Invasive prenatal diagnosis was performed by taking fetal cord blood, amniotic fluid, and fetal ascites respectively for cytogenetics, molecular genetics and biochemical examination and the impact of intrauterine therapeutic procedures on neonatal outcomes was evaluated as well. The correlation among prenatal examination, pathogeny and prognosis was analyzed by Fisher′s exact test.Results:(1) The prognosis of isolated fetal ascites initially presenting ≥28 weeks was better than that before 28 weeks, survival rate of 1-year-old were 13/15 and 9/17,respectively, the difference was statistically significant ( P<0.05). (2) The etiologic diagnosis rate of ascites before delivery was 31%(11/36), which increased to 53%(19/36) totally after birth. Characteristics of cases which were defined prenatally were as follows: 8 cases of digestive tract diseases showed ultrasonic abnormalities, including echogenic bowel, bowel dilatation and polyhydramnios; platelet level in umbilical cord blood of fetuses infected with cytomegalovirus were below 100 × 10 9/L in 2 cases; 1 case of urinary system malformation showed megalocystis and hydronephrosis. Cases which were defined causes after birth included: 3 fetuses with chyloperitonium presented persistent fetal ascites; 3 cases of digestive-related causes were rectal duplication with infection, mesentery stenosis, and intestinal atresia; other causes included Pierre-Robin syndrome and Budd-Chiari syndrome. (3) The live birth rate was 72% (26/36) and survival rate of 1-year-old was 61% (22/36). And 9/10 of infants who underwent surgeries got good outcomes. Fetal ascites due to abdominal or pelvic factors turned well in 13/16 of cases. Conclusions:The pregnancy outcome of fetal isolated ascites depends mainly on primary causes. Gastrointestinal abnormality is one of the most common causes. Excluded intrauterine infection, chromosomal abnormality and abnormal systemic ultrasonic findings, fetus with reduced ascites as the pregnancy progresses will get good outcome.

Newborn screening by tandem mass spectrometry in Nanjing: a retrospective analysis of 175 767 cases / 中华围产医学杂志

Objective:To analyze the detection of neonatal inherited metabolic diseases in Nanjing.Methods:We researched the results of 175 767 newborns by tandem mass spectrometry from December 2013 to July 2018. Amino acids, acylcarnitines, and succinylacetone were detected by non-derivatized tandem mass spectrometry to screen the abnormity of newborn amino acid, organic acid, or fatty acid oxidation metabolism disease. Gene panels based on high throughput sequencing technology were carried out to detect gene mutation of positive neonates. Descriptive statistics were used to analyze all the data.Results:The positive rate of primary screening was 2.1% (3 691/175 767), 3 598 of 3 691 positive cases were recalled. At last, 62 cases of the inherited metabolic disease were diagnosed. Among them, there were 35 cases of amino acid metabolism disease, 12 cases of organic acid metabolism disorder, and 15 cases of fatty acid metabolism defect. The total incidence of neonatal inherited metabolic disease was 0.035 3%, among which amino acid metabolic diseases were 0.019 9%, organic acid metabolic diseases were 0.006 8%, and fatty acid metabolic diseases were 0.008 5%. The diseases with the highest incidence were phenylalanine hydroxylase deficiency (0.015 9%), methylmalonic acidemia (0.005 1%), and primary carnitine deficiency (0.005 1%). Among 62 children, 51 (82.2%) were diagnosed by gene diagnosis (including 17 cases of phenylalanine hydroxylase deficiency and 34 cases of other inherited metabolic diseases). Another 11 children with phenylalanine hydroxylase deficiency refused gene diagnosis. Two pathogenic mutations were found in 17 children with phenylalanine hydroxylase deficiency. Two pathogenic mutations were found in 29 of the other 34 children with inherited metabolic disease, which were from their parents, while only one pathogenic mutation was found in the other five children, of which two cases with hypermethioninemia were autosomal dominant inheritance.Conclusions:Neonatal inherited metabolic diseases with high incidence in Nanjing are phenylalanine hydroxylase deficiency, methylmalonic acidemia, and primary carnitine deficiency. Some cases screened by tandem mass spectrometry only showed abnormal screening indicators. No specific clinical symptoms were found during follow-up, and further follow-up was needed.

Correlation analysis between HCY,LP-PLA2,RDW,and UA in ischemic stroke patients with H-type hypertension / 中风与神经疾病杂志

@#Objective　To explore the changes of HCY and LP-PLA2,RDW,and UA in patients with H-type hypertension with acute cerebral infarction.Methods　A total of 408 patients with acute cerebral infarction treated in the Department of Neurology,First Hospital of Lanzhou University from January 2017 to June 2019 were selected and divided into H type hypertension group (248 cases),HCY alone group (107 cases).The non-H hypertension group (31 cases),the simple acute cerebral infarction group (22 cases),and the healthy physical examination patients in the same period were selected as the control group (63 cases).Results　(1)The levels of male,LP-PLA2,HCY,and RDW were significantly higher in the H-type hypertension group than in the simple acute cerebral infarction group and the normal control group(P<0.05);(2)HCY levels were significantly higher in men with H-type hypertension than women(P<0.05);(3)HCY was positively correlated with RDW and UA in the H-type hypertension group (r=0.389,P<0.001;r=0.171,P<0.001);(4)In the H-type hypertension group,HCY,LP-PLA2,RDW,and UA all have certain diagnostic efficacy (AUC=0.724,0.916,0.731),respectively,0.677 were less than the joint detection AUC=0.947.Conclusion　HCY,LP-PLA2,RDW,and UA can prompt the inflammation status of H-type hypertension with acute cerebral infarction;H-type hypertension and LP-PLA2 levels have certain synergistic effect in acute cerebral infarction.

Analysis of AGR1 gene variants in an infant with early-onset argininemia / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for an infant with early-onset argininemia.@*METHODS@#Potential variant was detected with an Ion Torrent semiconductor sequencer using a gene panel for inherited diseases. Suspected variants were verified by Sanger sequencing.@*RESULTS@#Genetic testing indicated that he has carried c.560+2T>C and c.811T>C compound heterozygous variant of the AGR1 gene, which were inherited from his father and mother, respectively. Among these, c.560+2T>C was suspected to be pathogenic, while c.811T>C was of unknown clinical significance, and both were not reported previously.@*CONCLUSION@#The c.560+2T>C and c.811T>C compound heterozygous variants of the AGR1 gene probably underlies the argininemia in this child. Above finding has enriched the variant spectrum of the AGR1 gene.

Clinical and genetic analysis of two children suspected for argininosuccinic aciduria / 中华医学遗传学杂志

OBJECTIVE@#To analyze the clinical and genetic features of two children suspected for arginylsuccinuria aciduria.@*METHODS@#The patients were subjected to high-throughput sequencing using a gene panel.@*RESULTS@#Both patients had high citrulline (87.37-156.10 μmol/L) measured by mass spectrometry/mass spectrometry (MS/MS) upon neonatal screening but had no symptoms. Two compound heterozygous variants of the ASL gene were detected in patient 1 (exon 6: c.467C>T inherited from her father and exon 7: c.556C>T inherited from her mother), among which c.556C>T is novel. Patient 2 had mental retardation and two full siblings who had died of hyperammonemia. Two compound heterozygosity variants of the ASL gene were detected (exon 3: c.281G>T inherited from his father and intron: c.208-15T>A inherited from his mother). Both were novel mutations.@*CONCLUSION@#Variants of the ASL gene probably underlie the argininosuccinic aciduria in the two patients. Above findings have enriched the spectrum of ASL mutations.

Clinical application for pregnacy outcomes of radiofrequency ablation in complex multiple pregnancies / 中华妇产科杂志

Objective@#To investigate the safety and effectiveness of radiofrequency ablation (RFA) for selective fetal reduction in complex multiple pregnancies and analyze factors affecting perinatal outcomes.@*Methods@#This was a retrospective case series of 156 patients undergoing selective fetal reduction by RFA in Provincial Hospital Affiliated to Shandong University from July 22th, 2011 to September 12th, 2018. They were divided into five groups according to surgical indications, including 46 cases in the monochorionic twins discordant for fetal anomalies group, 42 cases in the multiple pregnancies for reducing fetal numbers group, 40 cases in the twin to twin transfusion syndrome (TTTS) group, 24 cases in the selective intrauterine growth restriction (sIUGR) group and 4 cases in the twin reversed arterial perfusion sequence (TRAPS) group. According to the gestational age at surgery, patients were divided into two groups: the gestational age at surgery <20 weeks group (75 cases) and the gestational age at surgery≥20 weeks group (81 cases). According to the cycles of RFA required for surgery, patients were divided into two groups: one cycle group (124 cases) and ≥2 cycles group (32 cases). Basic information of patients, surgical process, postoperative complications and pregnancy outcomes were recorded. The growth and development of survival newborns were also followed up. Factors affecting perinatal outcomes were analyzed.@*Results@#(1) The median gestational age at procedure of 156 patients was 20 weeks (14+5- 29+1 weeks). The median cycles of RFA was 1 cycle (1-3 cycles), of which one cycle accounted for 79.5% (124/156). (2) Eleven (7.1%, 11/156) patients experienced intrauterine fetal death, 27 (17.3%, 27/156) patients miscarried, and the overall survival rate was 75.6% (118/156). Premature birth rate before 34 weeks was 19.5% (23/118). There were 129 neonates. The median gestational age at delivery was 37+3 weeks (28+2- 41+1 weeks) with a mean birth weight of (2 657±700) g. (3) Analysis of pregnancy outcomes based on surgical indications found that, the gestational age at delivery [38 weeks (30+1-41+1 weeks), 36+4 weeks (29- 39 weeks), 36+4 weeks (28+2-39+5 weeks), 38 weeks (31-39+6 weeks), 38+3 weeks (30+4-38+4 weeks)] and neonatal birth weight [(2 820±671), (2 435±416), (2 497±843), (2 998±718), (2 517±1 087) g] were significantly different among fetal anomalies group, reducing fetal numbers group, TTTS group, sIUGR group and TRAPS group, respectively (all P<0.05). There were no significant differences in the pregnancy outcomes between gestational age at surgery <20 weeks group and gestational age at surgery ≥20 weeks group, or between one cycle group and ≥2 cycles group, respectively (all P>0.05).@*Conclusions@#RFA is a safe and effective procedure in treating complex monochorionic multiple pregnancies. Surgical indications would affect the gestational age at delivery and neonatal outcomes.

Intron mutations of argininosuccinate lyase (ASL) gene exist in a family with argininosuccinic aciduria / 临床检验杂志

Objective@#To perform gene detection and gene mutation analysis in a family with inherited metabolic diseases characterized as increased citrulline (Cit) by the MS/MS assay. @*Methods@#The peripheral blood samples were collected from the family members, and genomic DNA was extracted for gene diagnosis, which was performed by the whole exon sequencing method. The novel mutation gene was cloned into pcDNA3.1(+) vector, and its pathogenicity was verified by the Mini-gene assay in cultured cells in vitro. @*Results@#The clinical diagnosis of the proband as argininosuccinic aciduria (ASA) was clear. Two pathogenic mutations, c.281G＞T (p.Arg94Leu) and c.208-15 T＞A, were detected in the argininosuccinate lyase (ASL) gene, and they were not reported previously. The Mini-gene expression in vitro confirmed that c.208-15 T＞A could cause aberrant splicing, resulting in the retention of 13 bp in intron 2. @*Conclusion@#Two new pathogenic mutations of ASL gene, c.208-15 T＞A and c.281G＞T, are found in an ASA family, which enriches the mutation profile of ASL gene. The Mini-gene assay is a simple and effective tool for the research of intron mutations.

Analysis of AGR1 gene variants in an infant with early-onset argininemia / 中华医学遗传学杂志

Objective@#To explore the genetic basis for an infant with early-onset argininemia.@*Methods@#Potential variant was detected with an Ion Torrent semiconductor sequencer using a gene panel for inherited diseases. Suspected variants were verified by Sanger sequencing.@*Results@#Genetic testing indicated that he has carried c. 560+ 2T>C and c. 811T>C compound heterozygous variant of the AGR1 gene, which were inherited from his father and mother, respectively. Among these, c. 560+ 2T>C was suspected to be pathogenic, while c. 811T>C was of unknown clinical significance, and both were not reported previously.@*Conclusion@#The c. 560+ 2T>C and c. 811T>C compound heterozygous variants of the AGR1 gene probably underlies the argininemia in this child. Above finding has enriched the variant spectrum of the AGR1 gene.

Clinical features of patients with Becker muscular dystrophy and deletions of the rod domain of dystrophin gene / 中华医学遗传学杂志

OBJECTIVE To explore the clinical features of patients carrying deletions of the rod domain of the dystrophin gene. METHODS Clinical data of 12 Chinese patients with Becker muscular dystrophy (BMD) and such deletions was reviewed. RESULTS Most patients complained of muscle weakness of lower limbs. Two patients had muscle cramps, one had increased creatine kinase (CK) level, and one had dilated cardiomyopathy. CONCLUSION Compared with DMD, the clinical features of BMD are much more variable, particularly for those carrying deletions of the rod domain of the dystrophin gene. Muscular weakness may not be the sole complaint of BMD. The diagnosis of BMD cannot be excluded by moderately elevated CK. For male patients with dilated cardiomyopathy, the possibility of BMD should be considered.

Combined application of MS/MS and LC-MS/MS in the newborn screening of children with methylmalonic acidemia / 临床检验杂志

Objective To investigate the combined application value of tandem mass spectrometry ( MS/MS) and high performance liquid chromatogram-tandem mass spectrometry ( LC-MS/MS ) in the newborn screening of children with methylmalonic acidemia ( MMA ) . Methods The dried blood spot samples from the newborn with abnormal propionylcarnitine ( C3) or C3/acetylcarnitine ( C2) or C3/free carnitine ( C0) levels in the preliminary screening of MS/MS were collected, and the concentrations of MMA, methylcitric acid ( MCA) and homocysteine ( Hcy) in these samples were detected with LC-MS/MS. The neonates with increased MMA, MCA or Hcy levels were recalled, and their urinary organic acids were analyzed with gas chromatographic mass spectrometry (GC/MS). Last, gene mutation anal-ysis was performed to make a definite diagnosis.Results A total of 423 samples with abnormal C3 or C3/C2 or C3/C0 levels in the new-born screening were collected, and the positive rate of preliminary screening was about 1％. The LC-MS/MS results showed that 8 neo-nates had higher MMA and tHcy levels. The GC/MS results further showed that the level of MMA increased slightly. Conclusion The combined application of MS/MS and LC-MS/MS may increase the positive predictive value and decrease the false positive rate of MMA screening in the newborn, which may have an important clinical significane in the screening of inherited metabolic disorders.

Safety and efficacy of radiofrequency fetal ablation for monozygotic triplet and quadruplet pregnancies / 中华围产医学杂志

Objective To investigate the safety and efficacy of radiofrequency fetal ablation (RFA) in the treatment of monozygotic triplet and quadruplet pregnancies. Methods We analyzed retrospectively the clinical data of eight gravidas, including seven monozygotic triplets and one monozygotic quadruplets admitted to the Department of Obstetrics, Shandong Provincial Hospital Affiliated to Shandong University from March 2014 to January 2017. All of the eight women accepted ultrasound-guided selective fetal reduction by RFA to reduce to twins. Descriptive methods were used to analyze the perioperative status of the gravidas, maternal and fetal outcomes and neonatal follow-up. Results (1) In seven cases, the fetuses were deprived of blood flow after one heating cycle of radiofrequency ablation, while in the other, blood flow was stopped after two heating cycles. Heart beats of the reduced fetuses slowed down gradually after RFA, and stopped at 10, 20-25 and 40 minutes after operation in one, four and three cases, respectively. The conserved fetus showed normal heartbeats. (2) All patients accepted regular obstetrical examination after RFA. One was diagnosed with gestational diabetes mellitus at 26 weeks, and hospitalized for 4 d because of preterm labor at 30+6weeks. One women was hospitalized to receive a two-week tocolysis treatment one day after surgery, and diagnosed with severe preeclampsia at 35 weeks. One patient who had a fever six days after surgery and was hospitalized for antiinfection treatment progressed to inevitable abortion on the day of admission. The other five pregnant women had no abnormalities. (3) Except for one miscarriage, the rest seven cases all continued the pregnancy until delivery by cesarean, among which two with preterm premature rupture of membranes eventually delivered before term (35+1and 33 weeks), one with severe preeclampsia also preterm delivered (35+4weeks) and four term deliveries. Apgar scores at 1 and 5 minutes of all newborns were over 7. Three of the 14 newborns were hospitalized and recovered, including one pathological jaundice, one laryngeal stridor and one premature. The last follow-up in September 2018 of all 14 babies did not show any abnormalities. Conclusions RFA is a feasible treatment for monozygotic triplets and quadruplets.

Effects of Kaixin Jieyu Pills on monoamines and BDNF in rats with vascular depression / 中成药

AIM To aim at investigating the function of Kaixin Jieyu Pills (Ginseng Radix et Rhizoma,Bupleuri Radix,Morindae officinalis Radix,etc.) to vascular depression and their mechanism of action.METHODS A vascular depression model of rat was established by chronic unpredictable mild stress and separation after ligation of the bilateral common carotid arteries.The rats were treated with Kaixin Jieyu Pills and fluoxetine hydrochloride (FLU) for 28 days,respectively.The sucrose preference test and open-field test were performed.Cerebral perfusion was investigated with a Perfusion Speckle Imager.The monoamines including serotonin,dopamine and noradrenaline,and brain-derived neurotrophic factor (BDNF) were determined with ELISA and Western bolt,respectively.RESULTS Kaixin Jieyu Pills significantly increased sucrose preference,moving distance and cortical blood flow.Kaixin Jieyu Pills could upregulate,to different extents,the expressions of monoamines,including serotonin,noradrenaline and dopamine,and BDNF.Kaixin Jieyu Pills had a function similar to FLU in behavior,upregulating monoamines and BDNF,but it is superior to FLU in cortical blood flow.CONCLUSION Kaixin Jieyu Pills has the advantage of reducing the depression-like behavior and improving cerebral hypoperfusion,which might be mediated by the upregulation of the serotonin,noradrenaline and BDNF.