Treatment and follow-up of 82 children with propionic acidemia / 中华围产医学杂志

Objective:To investigate the treatment and prognosis of children with propionic acidemia (PA).Methods:This study involved 82 children with PA treated in the Department of Pediatric Endocrinol-ogy and Genetic Metabolism, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from December 2002 to June 2020. Clinical data, including manifestations, laboratory test results, treatment strategy, and follow-up data, were summarized and analyzed using t-test or Mann-Whitney U test. Results:(1) Among the 82 cases consisting of 50 (61.0%) boys and 32 (39.0%) girls, 59 (72.0%) were diagnosed after clinical onset; 22 (26.8%) were diagnosed by newborn screening, including eight asymptomatic ones; the other one (1.2%) was asymptomatic but confirmed after the diagnosis of PA in the patient's sibling. The average age at first onset was 4.5 months (2 d-5 years) in 73 subjects, of which 28 (38.4%) were early-onset PA (within three months after birth). (2) Cranial MRI was performed on 26 cases, and abnormality was identified in 19 (73.1%) cases. (3) Hyperlactatemia was found in 16 cases among 30(53.3%) who underwent relevant examination with the average lactic acid level of 3.5 (2.1-4.3) μmol/L, while 35 out of 40 patients (87.5%) had hyperammonemia with an average blood ammonia level of 105.4 (34-907) μmol/L. (4) Among the 28 early-onset PA cases, 16 (57.1%) died, and 12 (42.9%) survived. There was no significant difference in the serum propionylcarnitine level, propionylcarnitine to acetylcarnitine ratio, urine 3-hydroxypropionic acid, or methylcitrate level between the survival and death cases. (5) Genetic mutations were detected in 75 patients (91.5%), among which 26 (34.7%) carried PCCA gene mutations and 48 (64%) with PCCB gene mutations. One patient (1.3%) harbored one known pathogenic mutation in each of the PCCA and PCCB genes. All mutations were inherited from the parents. (6) Followed up to June 2020, 57 (69.5%) patients survived, and 25 (30.5%) died from multiple organ failure secondary to severe acidosis, including 16 early-onset and nine late-onset cases. Conclusions:The primary treatment of PA is dietary control. Most PA patients are diagnosed after clinical onset, but symptoms may recur and even have developmental retardation despite treatment. Some of those diagnosed through newborn screening are asymptomatic after treatment. Newborn screening using tandem mass spectrometry is recommended for early diagnosis and treatment of PA.

Retrospective analysis on clinical data and genetic variations of patients with beta-ketothiolase deficiency / 中华医学遗传学杂志

OBJECTIVE@#To summarize the clinical, biochemical and molecular characteristics of 8 patients with beta-ketothiolase deficiency (BKD).@*METHODS@#Clinical characteristics, biochemical markers detected by tandem mass spectrometry (MS-MS) and gas chromatography-mass spectrometry (GC-MS), and variations of ACAT1 gene of the 8 patients were reviewed.@*RESULTS@#Three patients were diagnosed by newborn screening and were asymptomatic. Five patients showed dyspnea and metabolic acidosis through high risk screening. Blood methylcrotonyl carnitine (C5:1) were 0.43 (0.20-0.89) μmol/L and 3-hydroxyisovaleryl carnitine(C5-OH) were 1.37 (0.98-3.40) μmol/L. Both were significantly higher than those of healthy controls (PG (p.N375S) variant, which accounted for 28.6% of all 14 mutant alleles. Four novel variants, namely c.229delG (p.E77KfsTer10), c.373G>T (p.V125F), c.419T>G (p.L140R) and c.72+1G>A, were discovered. Pathogenicity assessment of two highly conservative missense variants (p.V125F) and (p.L140R) were 0.994 and 1.0 (Scores obtained from PolyPhen2), and PROVEAN scores were -4.652 and -5.399, respectively. c.72+1g>a was suspected (by Human Splicing Finder) to alter the wild type donor motif and most probably affect the splicing.@*CONCLUSION@#Clinicians should consider MS/MS and GC/MS testing for those with unexplained neurological symptoms and metabolic acidosis in order to attain early diagnosis of BKD. Genetic testing should be used to confirm the diagnosis.

Clinical value of serum new molecular markers in the early diagnosis of sepsis in elderly patients with burns / 中华老年医学杂志

Objective@#To investigate the clinical value of the serum new molecular markers, soluble triggering receptor expressed on myeloid cells-1(sTREM-1)and soluble hemoglobin scavenger receptor(sCD163), in the diagnosis of sepsis in elderly patients with burns.@*Methods@#A total of 58 inpatients with burns from Jun 2017 to June 2018 were enrolled in the study.Patients were divided into three groups: the sepsis group(n=12), the localized infection group(n=21)and the non-infection group(n=29). The levels of sTREM-1 and sCD163 were determined by enzyme-linked immunosorbent assays(ELISAs). The clinical diagnostic value of sTREM-1 and sCD163 was assessed by receiver operating characteristic(ROC)curve analysis.@*Results@#There was a statistically significant difference in the levels of sTREM-1 and sCD163 at day 1 between the three groups(F=20.994 and 38.363, P<0.01). Serum levels of sTREM-1 and sCD163 were higher in the sepsis group than in the localized infection group and the non-infection group.Serum levels of sTREM-1 and sCD163 were higher in the localized infection group than in the non-infection group.Serum levels of sTREM-1 and sCD163 were lower at day 7 than those at day 1 in all groups(F=21.242 and 41.035, P<0.01). Serum sTREM-1 levels were positively correlated with serum sCD163 levels(r=0.609, P=0.000). The AUC of sTREM-1 and sCD163 for the diagnosis of sepsis was 0.880(95%CI: 0.816~0.926).@*Conclusions@#Serum levels of sTREM-1 and sCD163 are elevated with increasing degrees of infection.Monitoring serum sTREM-1 and sCD163 levels is helpful for the diagnosis of sepsis in elderly patients with burns.

Clinical value of serum new molecular markers in the early diagnosis of sepsis in elderly patients with burns / 中华老年医学杂志

Objective To investigate the clinical value of the serum new molecular markers,soluble triggering receptor expressed on myeloid cells-1 (sTREM-1)and soluble hemoglobin scavenger receptor(sCD163),in the diagnosis of sepsis in elderly patients with burns.Methods A total of 58 inpatients with burns from Jun 2017 to June 2018 were enrolled in the study.Patients were divided into three groups:the sepsis group(n=12),the localized infection group(n=21)and the non-infection group (n=29).The levels of sTREM-1 and sCD163 were determined by enzyme-linked immunosorbent assays(ELISAs).The clinical diagnostic value of sTREM-1 and sCD163 was assessed by receiver operating characteristic(ROC)curve analysis.Results There was a statistically significant difference in thelevels of sTREM-1 and sCD163 at day 1 between the three groups(F =20.994 and 38.363,P＜0.01).Serum levels of sTREM-1 and sCD163 were higher in the sepsis group than in the localized infection group and the non-infection group.Serum levels of sTREM-1 and sCD163 were higher in the localized infection group than in the non-infection group.Serum levels of sTREM-1 and sCD163 were lower at day 7 than those at day 1 in all groups(F=21.242 and 41.035,P＜0.01).Serum sTREM-1 levels were positively correlated with serum sCD163 levels (r =0.609,P =0.000).The AUC of sTREM-1 and sCD163 for the diagnosis of sepsis was 0.880(95％CI:0.816～0.926).Conclusions Serum levels of sTREM-1 and sCD163 are elevated with increasing degrees of infection.Monitoring serum sTREM-1 and sCD163 levels is helpful for the diagnosis of sepsis in elderly patients with burns.

Clinical analysis and outcome of patients with c. 482G >A variant of MMACHC gene in cblC type methylmalonic acidemia / 中华内分泌代谢杂志

Objective To explore the clinical manifestations, treatment and outcomes of patients with c. 482G>A ( p. R161Q ) variant of MMACHC gene in cblC type methylmalonic acidemia ( MMA ) . Methods The clinical manifestations, mass spectrometry results, genotypes, treatment and outcomes of 75 patients with cblC type MMAcarryingc.482G>A(p.R161Q)variantwereretrospectivelyanalyzed.Results Ofthe75patients,57(76％) were from newborn screening and one of them had an onset. Among the rest 18 unscreened patients, 2 were diagnosed after their full sisters' or brothers' diagnosis, the others were clinical patients. There were 17 clinical patients, with the medium age of onset 12 years old (10 days～26 years old). 12 late onset patients (70.6％) presented with poor academic performance, memory loss, poor expression, and decreased exercise capacity, while 5 early onset patients (29.4％) presented with convulsion and delay of development. All patients were vitamin B12-responsive. The levels of blood propionylcarnitine, the ratio of propionylcarnitine to acetylcarnitine, urinary methylmalonic acid and methyldecanoic acid, and plasma homocysteine were significantly decreased after treatment (P< 0.01). All patients diagnosed from newborn screening had normal development. However, only 3 clinical patients had a rather normal outcomes and the others remained different levels of intelligence and ( or ) motor dysfunction after treatment. Conclusion The c.482G>A ( p. R161Q) variant of MMACHC gene is associated with late onset cblC type MMA. Patients with this variant have a better response to hydroxycobalamin than other variants. The outcome of patients diagnosed from the newborn screening is good. When symptoms occur, the disability rate is often high. Therefore, newborn screening is a recommended method to prevent this disease.

Mass spectrometry combined with gene analysis for prenatal diagnosis of glutaric acidemia type Ⅰ / 中华儿科杂志

Objective@#To investigate the value of amniotic fluid metabolite detection by mass spectrometry combined with gene mutation analysis in the prenatal diagnosis of glutaric acidemia type Ⅰ (GA-Ⅰ).@*Method@#From January 2009 to December 2016, Department of Pediatric Endocrinology and Genetic, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine carried out prenatal diagnosis for 24 cases of pregnant women with GA-Ⅰproband. 24 pregnant women without organic acidemia proband for conventional prenatal diagnosis at the same period were used as the control group. The pregnant women of the two groups had the amniocentesis at 16 to 20 weeks of gestation.The levels of glutaryl carnitine (C5DC) and octanoylcarnitine (C8) in amniotic fluid were detected by tandem mass spectrometry, and the levels of glutaric acid was determined by gas chromatography-mass spectrometry. All the amniotic fluid cells underwent GCDH gene testing.@*Result@#A total of 4 cases of fetuses were diagnosed by gene mutation analysis combined with mass spectrometry detection, the levels of C5DC (1.58(0.89-2.85) μmol/L), C5DC/C8 (19.74(12.40-25.93))and glutaric acid (129.96 (90.09-66.02) mmol/mol Cr) were significantly higher than the upper limit of the reference, of which in one case with the proband only on mutation was detected, and in the amniotic fluid cells also only one mutation was detected, the diagnosis was made according to the significantly increased levels of amniotic fluid C5DC, C5DC/C8 and glutaric acid. Twenty cases of fetuses were identified as non-GA-Ⅰchildren, of whom in 2 cases of proband only one mutation was detected, and also in amniotic fluid cells one mutation was detected, in 2 cases the diagnosis was excluded because the normal levels of C5DC, C5DC/C8 and glutaric acid. There were 2 cases whose levels of C5DC or glutaric acid were slightly higher than the upper limit of the reference, but the diagnosis was excluded according to genetic testing.@*Conclusion@#Prenatal diagnosis cannot be made by gene analysis when the proband mutation is not clear, and it cannot determine whether the fetus is patient when the mass spectrometry detection of amniotic fluid metabolite is mildly abnormal, while mass spectrometry detection of amniotic fluid C5DC, C5DC/C8 and glutaric acid levels combined with GCDH gene analysis can make up the deficiencies, and make the prenatal diagnosis of GA-Ⅰ more reliably.

Prenatal diagnosis of methylmalonic acidemia using test of homocysteine and mass spectrometry in amniotic fluid / 中华检验医学杂志

Objective To explore the value of the combination of homocysteine analysis, liquid chromatography tandem mass spectrometry(LC-MS/MS)and gas chromatography mass spectrometry(GC/MS)in the prenatal diagnosis of combined methylmalonic acidemia and homocystinuria(cblC defect)in amniotic fluid.Methods This is a retrospective study of 187 cases of pregnancies that came to our hospital for prenatal diagnosis between 2014/01-2017/03,among which 78 cases′probands were cblC defect patients and 109 cases′probands were not organic academia patients(control group).Amniotic fluid samples from pregnant women were obtained at 16 -24 weeks of gestation.Propionylcarnitine(C3)and acetylcarnitine (C2)were measured by LC-MS/MS, methylmalonic acid and methylcitric acid were analyzed by GC /MS, and homocysteine was determined by fluorescence polarization immunoassay.Some pregnancies received MMACHC gene sequencing with cultured cells from amniotic fluid.Data were analyzed using Mann-Whitney U and Kruskal-Wallis H tests.Results Among those 78 pregnant women whose probands were diagnosed to be cblC defect,24 cases were diagnosed to be cblC defect(positive group)and 54 pregnant women were diagnosed to be negative(negative group).In positive group, levels of homocysteine, C3, C3/C2, methylmalonic acid and methylcitric acid were all significantly higher than their normal reference ranges, negative group and control group(P values are 0.00).Cases that were diagnosed to be cblC defect by MMACHC gene sequencing were all turned out to be positive in the tests of the above metabolites in amniotic fluid.Cases with negative results of the metabolites were all excluded to be cblC defect by gene sequencing. Besides,2 cases of pregnancies were diagnosed to be positive by homocysteine and mass spectrometric analysis while only one mutation were detected by gene sequencing.Conclusions The combination of homocysteine, LC-MS/MS and GC/MS analysis in amniotic fluid turns out to be reliable for prenatal diagnosis of cblC defect,which may further cover the defect of prenatal diagnosis of those pregnancies whose probands′gene mutation is unknown.

Prenatal diagnosis of methylmalonic acidemia by amniotic fluid metabolites analysis using mass spectrometry / 中华围产医学杂志

Objective To investigate the effect of tandem mass spectrometry and gas chromatography-mass spectrometry to make prenatal diagnosis of methylmalonic acidemia (MMA) by detecting organic acid and acylcarnitine in amniotic fluid.Methods From October 11,2007 to December 20,2014,131 pregnant women with MMA proband received prenatal diagnosis of MMA in Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (case group).Another 120 cases of pregnant women for conventional prenatal diagnosis at the same period were as control group.The pregnant women of two groups had the amniocentesis at 16 to 20 weeks of gestation.The levels of propionylcarnitine(C3) and acetylcarnitine(C2)in amniotic fluid were detected by tandem mass spectrometry.The methylmalonic acid and methylcitrate acid were detected by gas chromatography-mass spectrometry.MMA gene of cells in amniotic fluid of eighty fetuses with proband clearly diagnosed were detected by gene testing.Data were analyzed by Wilcoxon test.Results In case group,29 fetuses were found positive for higher level of C3,C3/C2,methylmalonic acid and methylcitrate acid compared with normal reference value,and the detected rate of fetal MMA was 22.1％(29/131).The levels of C3 and C3 / C2 in amniotic fluid of these 29 cases were higher than those in control group[8.13(2.42-16.70) vs 1.04(0.52-3.40) μmol/L,Z =-8.313; 0.77(0.30-1.79) vs 0.10(0.05-0.22),Z=-8.374; P ＜ 0.05 respectively].The levels of methylmalonic acid and methylcitrate acid were also higher[9.13(1.68-61.78) vs 0.00(0.00-1.31) mmol/mol Crea,Z=-11.348; 0.58(0.00-1.90) vs 0.05(0.00-0.52) mmol/mol Crea,Z=-6.632,P ＜ 0.05 respectively].For the other 102 cases in case group,the levels of C3,C3/C2,methylmalonic acid and methylcitrate acid were not higher than normal reference value,and were similar to those in control group (P ＞ 0.05); while they were lower than those of positive MMA fetuses (all P ＜ 0.05).Among 29 positive fetuses,16 fetuses were detected MMA gene,five were diagnosed as MUT forms of MMA and 11 were MMACHC forms of MMA.In 102 MMA negative fetuses,64 fetuses were detected MMA gene,44 were found one mutant site and 20 were found no gene mutation.The coincidence rate between gene detecting and mass spectrometry was 100％(80/80).Conclusions Mass spectrometry could be used to measure the C3,methylmalonic acid and methylcitrate acid levels in amniotic fluid of pregnant women with MMA proband to make prenatal diagnosis.

Progress on diabetic cognitive dysfunction / 基础医学与临床

Diabetic cognitive dysfunction is one of the important complications of diabetes mellitus .Insulin plays an important protective role in cognitive function through MAPK and PI 3-K/Akt signaling pathway .Insulin resistance may give rise to excessive tau protein phosphorylation , formation of neurofibrillary tangles inhibits insulin degrading enzyme , reduces the degradation of amyloid β;enhances oxidative stress , further damages the integrity of the neu-ronal structure and function and eventually leads to cognitive dysfunction .

DTI study of corpus callosun for evaluating diffuse axonal injury / 实用放射学杂志

Objective To evaluate DTI for early diagnosis and prognosis of diffuse axonal injury (DAI)in the moderate and sever traumatic brain injury (sTBI).Methods 30 patients with sTBI and 30 healthy controls were studied with T1 WI and DTI.The frac-tional anisotropy(FA)and ADC were quantified from different regions of interest(ROI)including the genu of corpus callosum(gCC), the trunk of CC,the splenium of CC(sCC)and the integrity CC.The FA and ADC of the same R0I was compared between TBI group and control group,and FA and ADC of the different parts of CC were compared in TBI group.Correlations between the FA or ADC and coma duration of the TBI patients were analyzed by Pearson linear correlation.Results Compared with control group,the FA value decreased significantly in each ROI,while ADC was reversed.The FA or ADC of the different ROI were correlated with co-ma duration in TBI group.Conclusion DTI is sensitive to detect the white matter injury of TBI.FA value detected in the CC can help diagnose DAI earlier and evaluate the degree of injury.

Expression and significance of Toll like receptor 2 and Toll like receptor 4 in chronic rhinosinusitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the role of the innate immune factors TLR2 and TLR4 in the pathogenesis of chronic rhinosinusitis (CRS) by detecting their expression in different clinical types of CRS and the normal control group.@*METHOD@#Immunohistochemistry was used to detect the expression of TLR2 and TLR4 respectively in 21 cases (chronic rhinosinusitis with nasal polyps, CRSwNP) group, 15 cases (chronic rhinosinusitis without nasal polyos, CRSsNP) group, 11 cases recurrent CRSwNP group and 13 cases control group. Positive cells were counted under the microscope artificially, Mann-Whitney U analysis was applied for the ranked data, and one-way anova analysis was adopted to analyze the experimental group and control group.@*RESULT@#(1) TLR2 and TLR4 expression had the same characteristics. Expression mainly concentrated in parts of the whole layer of epithelial basement membrane, cytoplasm of glandular cells, very few inflammatory cells such as monocytes and plasma cells in the cytoplasm, sometimes unknown cell nuclei positive expression. (2) The glandular cells were stained manual counting and color grading. TLR2 and TLR4 packet application Wilcoxon rank test Mann-Whitney U test analysis was not statistically significant (P > 0.05), measurement data within the group variance statistical difference between the groups (P < 0.05).@*CONCLUSION@#The Nasal mucosa can produce the innate immune factors TLR2 and TLR4. The different expression of TLR2 and TLR4 in the various clinical types of CRS suggests that they play the certain role in the pathogenesis of CRS.

Expression of TGF-beta1 and collagen fibers in chronic nasal-sinusitis nasal mucosa of patients / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Explore the TGF-beta1 and collagen fibers in chronic nasal sinusitis each type and degree of expression of the normal control group and in the nasal mucosa epithelial tissue remodeling and the role of TGF-beta1 and collagen deposition relationship.@*METHOD@#Sixty-two patients experimental group (CRSwNP of 21 cases. CRSs NP group of 15 cases. 11 cases of recurrent nasal polyps; control group 15 cases specimens for immunohistochemistry and masson collagen staining. Manual counting the number of positive cells by the Mann-Whitney U test to analyze the expression in experimental group and the control group. Experimental group and control group between the single-factor analysis of variance with a One-Way ANOVA analysis.@*RESULT@#Experimental group and control group were expressed TGF-beta1 with collagen deposition. Which, TGF-beta1 in CRSsNP group was significantly lower than the control group (P < 0.05), CRSwNP group was significantly increased compared with CRSsNP group (P < 0.05); masson collagen staining, CRSsNP group was significantly lower than the control group (P < 0.01), recurrent nasal polyps group than in the control group was significantly increased (P < 0.05). TGF-beta1 and collagen staining masson positive correlation between (P < 0.01 ).@*CONCLUSION@#TGF-beta1 and collagen deposi tion and chronic nasal-sinusitis tissue remodeling has its relevance. Furthermore. TGF-beta1 expression increased with excessive deposition of collagen fibers also positively correlated.