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Objective:To investigate the accuracy, effectiveness and feasibility of MassARRAY genotyping assay in the diagnoses of neonatal genetic metabolic diseases.Methods:This is a retrospective study. From December 2016 to January 2020, newborns were screened by tandem mass spectrometry at the Zhejiang Newborn Screening Center, among which the data of 7 922 suspected positive cases of genetic metabolic diseases were collected. These patients were then tested for the common variants of 27 genetic metabolic diseases by MassARRAY genotyping assay, along with further testing using Sanger or next-generation sequencing used to verify and/or further search for potential variants.Results:A total of 1 408 cases were tested with MassARRAY. Among these, 307 cases were confirmed with certain genetic metabolic diseases. The detection rate of hyperphenylalaninemia was the highest, followed by primary carnitine deficiency, short acyl-coA dehydrogenase deficiency and methylmalonic acidemia. With these cases, the consistency of Sanger sequencing and MassARRAY was 100% (307/307). Another 287 cases were identified as carriers by MassARRAY with a 49.1% (141/287) consistency in reference to Sanger sequencing, mainly involving SLC22A5 and MCCC1 genes. Meanwhile, 50.8% (146/287) of these cases were found to have another variant mainly involving PAH, PTS and ACADS genes. The remaining 814 cases have no variants; 158 cases out of these patients have continuously abnormal amino acids, acyl carnitines, urine organic acid and/or other biochemical indices, and were tested by next-generation sequencing, among which 38% (60/158) were detected with two variants. In this study, a total of 513 patients with genetic metabolic disease were diagnosed, and the detection rate of MassARRAY was 59.8% (307/513). Conclusions:MassARRAY genotyping assay can be used as an early molecular screening method for neonatal genetic metabolic diseases. The detection rate is particularly high in diseases with a high concentration of hotspot variants, such as hyperphenylalaninemia and primary carnitine deficiency. The future application value of MassARRAY should be further improved by continuously optimizing its ability to identify new disease genes and potential variable sites.
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Objective:To investigate the incidence, clinical characteristics and prognosis of ornithine transcarbamylase deficiency(OCTD) in newborns in Zhejiang Province.Methods:A retrospective research was conducted.A total of 4 261 036 newborns from Department of Genetics and Metabolism, Children′s Hospital, Zhejiang University School of Medicine, between January 2009 and December 2021 were screened for inherited metabolic disorders using tandem mass spectrometry.OCTD was confirmed by urine organic acid and OTC gene analysis.Patients with OTCD received guidance on diet and lifestyle management, and were treated with citrulline and arginine.Long-term follow-up was performed.Their growth and intellectual development were evaluated. Results:A total of 7 patients with OCTD were diagnosed, with an incidence of 1.6/1 million.All patients were males.Two patients had neonatal-onset OCTD, and the other 5 had late-onset OCTD.Symptoms occurred several times in 6 patients, inducing hyperammonemia and hepatic impairment.One patient had no clinical manifestation.One patient died in the neonatal period.Blood citrulline levels were decreased in 7 patients to varying degrees.Uracil levels were increased in 4 patients, and 1 of them was complicated with elevated orotic acid levels.All patients had hemizygote variations in the OTC gene, including 6 missense variations(c.604C>T, c.386G>A, c.779T>C, c.1019C>T, c.594C>G, c.931G>A) and 1 intron variation(c.514-35C>G). Two variants(c.594C>G, c.514-35C>G) were never reported previously. Conclusions:The OTCD incidence by newborn screening is low with 1.6/1 million in Zhejiang province.All patients are males and present hypocitrullinemia.The clinical manifestations of OTCD are highly heterogeneous.The neonatal-onset form is severe and survivors always suffer serious sequelae.The late-onset form is mostly manifested with hyperammonemia and hepatic impairment.There may be association between phenotype and genotype.Two novel OTC variants are identified, which further expands the mutational spectrum.
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Objective:To investigate the efficiency of biochemical screening and hotspot gene screening in the detection of neonatal inherited metabolic diseases.Methods:This was a prospective multi-center study.The study was carried out on 21 442 neonatal samples collected from 12 hospitals in 10 provinces from November 2020 to November 2021.The results of biochemical screening and hotspot gene screening were analyzed jointly.Biochemical screening methods included glucose-6-phosphate dehydrogenase deficiency enzyme activity assay and neonatal tandem mass spectrometry.Genetic screening analysis involved 135 genes associated with 75 neonatal diseases.Results:Of all the 21 442 neonates enrolled in the study, 21 205 were subject to biochemical screening.A total of 813 cases were positive in the initial screening, and 0.45% of them (95 cases) were diagnosed after recall.All the 21 442 neonates underwent gene screening.About 168 positive cases were detected in the initial screening, and 0.73% (156 cases) of them were confirmed finally.Biochemical and genetic screening improved the detection sensitivity of such diseases as primary carnitine deficiency, neonatal intrahepatic cholestasis caused by citrin deficiency, and 2-methylbutyrylglycinemia.Moreover, biochemical and genetic screening enabled the detection of more diseases, including the common single-gene genetic diseases such as thalassemia and Wilson disease.Conclusions:In neonatal screening, the combination of biochemical screening and gene screening expands the number of diseases detected and improve screening efficiency.
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OBJECTIVE@#To investigate the clinical manifestations, biochemical abnormalities and pathogenic variants among children with Short/branched-chain acyl-CoA dehydrogenase (SBCAD) deficiency detected by neonatal screening.@*METHODS@#A total of 2 730 852 newborns were screened from January 2016 to December 2021 with liquid chromatography tandem mass spectrometry. Suspected SBCAD deficiency patients were diagnosed by urine organic acid analysis and high-throughput gene sequencing analysis. The clinical, biochemical and genetic changes of the confirmed cases were analyzed, in addition with guidance for diet and life management, L-carnitine supplement, and survey of growth and intellectual development.@*RESULTS@#Twelve cases of SBCAD deficiency were diagnosed, which yielded a prevalence of 1/227 571. The lsovaleryl carnitine (C5) of primary screening blood samples was between 0.6 and 2.1 µmol/L, all exceeded the normal range. C5/acety1 carnitine (C2) was between 0.02 and 0.12, with 6 cases exceeding the normal range. C5/propionyl carnitine (C3) was between 0.1 and 1.16, with 5 cases exceeding the normal range. Free carnitine (C0) was between 18.89 and 58.12 µmol, with 1 case exceeding the normal range. Three neonates with abnormal screening results were recommended to have appropriate restriction for protein intake and two were given L-carnitine. During follow-up, their C5 has ranged from 0.22 to 2.32 µmol/L, C5/C2 has ranged from 0.01 to 0.31, C5/C3 has ranged from 0.14 to 1.7. C5 or C5/C2 and C5/C3 were transiently normal in all patients except for case 8 during the neonatal screening and follow-up. C0 was 17.42 ∼ 76.83 µmol/L Urine organic acid analysis was carried out in 9 of the 12 cases, and 2-methylbutyroglycine was elevated in 8 cases. Urine organic acid analysis was carried out in 9 cases, and 2-methylbutyrylglycine was increased in 8 cases. Genetic analysis was carried out for 11 children, and in total 6 ACADSB gene variants were identified, which included 4 missense variants (c.655G>A, c.923G>A, c.461G>A, c.1165A>G), 1 frameshift variant (c.746del) and 1 nonsense variant (c.275C>G). Among these, the C.461G>A variant was unreported previously. The most common variants were c.1165A>G (40.9%) and C.275C>G (22.7%). The patients were followed up for 18 days to 55 months. Only one patient had mental retardation, with the remainders having normal physical and mental development.@*CONCLUSION@#SBCAD deficiency is a rare disease. The detection rate of newborn screening in this study was 1/227 571. Early intervention can be attained in most asymptomatic patients through neonatal screening. In this study, the common gene variants are c.1165A>G and c.275C>G.
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Humans , Infant, Newborn , Amino Acid Metabolism, Inborn Errors/genetics , Carnitine , Neonatal Screening/methodsABSTRACT
Abstract@#Transport of filter paper-dried blood spot samples is a critical procedure during the screening of neonatal inherited metabolic diseases, which is of great significance for the screening accuracy. In order to ensure the timing and safety of sample transport, the cold chain positioning system was initiated by Zhejiang Provincial Center for Quality Control of Neonatal Disease Screening since March 2015. Based on the framework of neonatal disease screening information management system, the function of the logistics transport management system was included in this positioning system, with aims to achieve the monitoring and tracking of sample transport processes through real-time positioning of the sample transport box via China Unicom 4G logistics card and global positioning system/BeiDou Navigation Satellite System. The samples are maintained in a transport environment at 2 to 8 °C via the temperature-controlled box made of 5 °C phase-changed cold-stored materials and general packet radio service (GPRS) temperature recorders. The mean pretest turnover duration reduced from 8.44 days to 5.03 days following introduction of the cold chain positioning system, and the percentage of timely sample delivery increased from 31.69% to 77.90%, while the withdrawal rate of unqualified samples reduced from 0.12% to 0.08%. The cold chain positioning system meets the requirements of transport of filter paper-dried blood spot samples, which has a high potential in screening of neonatal inherited metabolic diseases.
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OBJECTIVE@#To explore the genetic basis for a child with mental retardation.@*METHODS@#The child was subjected to next generation sequencing (NGS). Candidate variant was analyzed with bioinformatic software.@*RESULTS@#NGS revealed that the child has carried a de novo heterozygous c.4035G>C (p.Gln1345His) variant of the ARID1B gene. The variant was unreported previously and may cause instability of the protein structure.@*CONCLUSION@#The de novo missense variant of ARID1B gene may underlie the mental retardation in the child. Above result has enabled genetic counseling and prenatal diagnosis for her family.
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OBJECTIVE@#To explore effects of different delivery and storage conditions on concentrations of amino acids and carnitines in neonatal dried blood spots (DBS), so as to provide evidence for improving accurate and reliable detection by tandem mass spectrometry.@*METHODS@#A total of 1 254 616 newborn DBS samples in Newborn Screening Center of Zhejiang Province were delivered and stored at room temperature (group A, @*RESULTS@#The concentrations of amino acids and carnitines in the three groups were skewed, and the differences in amino acid and carnitine concentrations among groups were statistically significant (all @*CONCLUSIONS@#Cold-chain logistics system and storage in low temperature and low humidity can effectively reduce degradation of some amino acids and carnitines in DBS, improve the accuracy and reliability of detection, and thus ensures the quality of screening for neonatal metabolic diseases.
Subject(s)
Humans , Infant, Newborn , Amino Acids/analysis , Carnitine/analysis , Dried Blood Spot Testing/standards , Humidity , Neonatal Screening , Reproducibility of Results , Specimen Handling/standards , Tandem Mass Spectrometry , Temperature , Time FactorsABSTRACT
OBJECTIVE@#To investigate the incidence,clinical,biochemical and genetic characteristics of isovaleric acidemia (IVA) in Zhejiang province.@*METHODS@#Between January 2009 and December 2019, a total of 3 510 004 newborns were screened for IVA using tandem mass spectrometry. Patients of IVA were confirmed by urine organic acid and @*RESULTS@#A total of 15 patients with IVA were diagnosed, with an incidence of 1/234 000. Three patients had acute neonatal IVA, and the rest were asymptomatic. The isovalerylcarnitine (C5) levels were increased in all patients. Twelve children underwent urinary organic acid analysis, of which 11 cases had elevated isovalerylglycine levels, 4 cases with 3-hydroxyisovalerate increased simultaneously. Eleven IVA patients underwent genetic testing, 9 patients were compound heterozygous variants in @*CONCLUSIONS@#The clinical manifestations of IVA are non-specific, and the gene spectrum is scattered. Newborn patients screened by tandem mass spectrometry can receive early diagnosis and treatment, so as to correct metabolic defects and pathophysiological changes.
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Child , Humans , Infant, Newborn , Amino Acid Metabolism, Inborn Errors/epidemiology , China/epidemiology , Isovaleryl-CoA Dehydrogenase/genetics , Mutation , Neonatal Screening , Tandem Mass SpectrometryABSTRACT
Objective:To investigate the application feasibility of Region 4 Stork (R4S) system, an international collaborative newborn screening data platform, combined with cut-off value analysis in the neonatal screening for very long chain acyl-CoA dehydrogenase deficiency (VLCADD) by tandem mass spectrometry (MS/MS).Methods:The retrospective study was performed in 2, 040 072 neonates screened by MS/MS in Neonatal Screening Center of Zhejiang Province, China from October 2013 to July 2018. Nine hundred and ten cases were determined and identified as suspected positive VLCADD neonates by traditional cut-off method of tandem mass spectrometry. The original data of these 910 screened neonates were further analyzed by R4S system. Based on clinical diagnosis and ACADL gene test results, the screening efficiency between two methods was statistically compared.Results:The data of 910 suspected VLCADD-positive cases interpreted by cut-off method were further analyzed by R4S system, and the positive interpretation was reduced to 238 cases (including 9 confirmed positive cases). A total of 16 different mutations were found in ACADL gene sequencing among the confirmed children. The screening false positive rate (FPR) declined from 0.44‰ (901/2 040 072) to 0.11‰ (229/2 040 072), the rate of positive predictive value (PPV) increased from 0.99% (9/910) to 3.78% (9/238), and the specificity increased from 99.96% (2 039 162/2 040 063) to 99.99% (2 039 834/2 040 063). There was a statistically significant difference between cut-off method alone and cut-off method combined R4S system analysis (χ2=393.5, P<0.05). Conclusions:The R4S system combined with cut-off method applied in VLCADD neonatal screening by MS/MS can effectively improve screening performance, reduce false positive rate, and has certain value in clinical application.
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OBJECTIVE@#To investigate the genetic characterization of 3-hydroxyisovalerylcarnitine (C5-OH) metabolic abnormality in neonates.@*METHODS@#Fifty two newborns with increased C5-OH, C5-OH/C3 and C5-OH/C8 detected by tandem mass spectrometry during neonatal screening were enrolled in the study. Genomic DNA was extracted from the whole blood samples of 52 cases and their parents. Seventy-nine genes associated with genetic and metabolic diseases including , were targeted by liquid capture technique. Variation information of these genes was examined by high-throughput sequencing and bioinformatic analysis, and then was classified based on the American College of Medical Genetics and Genomics (ACMG) standards and guidelines. The genetic types were classified as wild-type, -maternal-mutation, -paternal-mutation and -mutation. Wilcoxon rank-sum test was performed for the increased multiples of C5-OH calculated in neonatal screening.@*RESULTS@#Twenty one variants (14 novel) were identified in 37 cases, 6 variants (5 novel) in 4 cases. The increased multiple of C5-OH calculated in -maternal-mutation and -mutation groups were significantly higher than that in wild-type group (all 0.05).@*CONCLUSIONS@#Mutations on and genes are the major genetic causes for the increased C5-OH in neonates, and maternal single heterozygous mutation can contribute to the moderately to severely increased C5-OH.
Subject(s)
Female , Humans , Infant, Newborn , Male , Carbon-Carbon Ligases , Genetics , Carnitine , Metabolism , Genetic Testing , Genetic Variation , Mutation , Neonatal Screening , Urea Cycle Disorders, Inborn , GeneticsABSTRACT
Objective To investigate the feasibility of Region 4 Stork(R4S)project used for newborn screening by tandem mass spectrometry in China.Methods This retrospective study was performed among 362 822 neonates screened by tandem mass spectrometry from May 2015 to April 2016 in Zhejiang newborn screening center.Infants were grouped by screening result category: 83 true positive cases,360 554 true negative cases and 2 185 false positive cases.Raw data was uploaded into R4S website to perform postanalytical interpretive tools, then results were analyzed with interpretation rules.The comparisons of normal population percentiles were done at five selected percentiles between Zhejiang newborn screening center and R4S project with min-max normalization.Results Compared with cutoff system by using R4S project with interpretation rules,the positive predictive value increased from 3.7%to 8.3%,the specificity increased from 99.40%to 99.75%, and the false positive rate declined from 0.6% to 0.2%. The two cases of true positive hyperprolinemia were reported negative, and one case of β-ketothiolase deficiency was misdiagnosis.Totally 311 638 cases in true negative group were resolved by postanalytical interpretive tools,and the remaining 48 916 cases were excluded with interpretation rules.False positive cases were reduced to 897 cases.Results of percentiles comparison showed that levels of some markers were significantly different between zhejiang newborn screening center and R 4S project.Conclusions R4S project effectively improved the newborn screening performance, whereas leaded to a small number of misdiagnosis and missed diagnosis.Besides,many true negative cases should be excluded with interpretation rules.Optimization should be achieved based on local normal population.(Chin J Lab Med,2018,41:300-304)
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<p><b>OBJECTIVE</b>To analyze the result and follow-up data of screening for newborn amino acid metabolic disorders in Zhejiang province.</p><p><b>METHODS</b>A total of 1 861 262 newborns were screened for amino acid metabolic disorders during January 2009 and December 2016 in Zhejiang province. The screening results and the follow-up data were analyzed retrospectively.</p><p><b>RESULTS</b>One hundred and sixty four cases were diagnosed as amino acid metabolic disorders with a prevalence of 1:11 349, including 83 with hyperphenylalaninaemia (1:22 400), 29 with neonatal intrahepatic cholestasis caused by citrin deficiency (1:64 138), 16 with methionine S-adenosyltransferase deficiency (1:116 250), 9 with maple syrup urine disease (1:206 667), 8 with argininemia (1:232 500), 7 with citrullinemia type Ⅰ (1:265 700), 6 with hyperprolinemia type Ⅰ (1:310 000), and 2 with carbamylphosphate synthetase Ⅰ deficiency(1:930 000). In addition, ornithine transcarbamylase deficiency, cystathionine β-synthase deficiency, argininosucoinate aciduria and tyrosinemia type Ⅰ were detected in one patient for each, respectively. Two patients had developmental delay, 7 patients were dead, and 2 cases of maple syrup urine disease were lost to follow-up.</p><p><b>CONCLUSIONS</b>Hyperphenylalaninaemia is the most common amino acid metabolic disease in newborns in Zhejiang province. Patients with amino acid metabolic disorders identified in newborn screening program can have chance for normal growth development by intervention.</p>
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<p><b>OBJECTIVE</b>To analyze the results and follow up data of screening for newborn organic aciduria in Zhejiang province.</p><p><b>METHODS</b>The results and follow-up data of 1 861 262 newborns from Zhejiang province undergoing screening for organic aciduria during January 2009 and December 2016 were retrospectively analyzed. The acylcarnitine spectrum in urine samples was detected by tandem mass spectrum (MS/MS) and the positive patients were confirmed by urine gas chromatography mass spectrometry and/or gene analysis.</p><p><b>RESULTS</b>Ninety two cases of organic aciduria were confirmed with a prevalence of 1:20 200. Among 40 cases of methylmalonic academia (MMA), 13 (32.5%) were of MMA simple type and 27 (67.5%) were combined type. Genetic analysis showed 6 cases of MUT type and 1 case of CblB type out of 7 patients with MMA simple type, 10 cases of CblC and 1 case of CblF out of 11 patients with combined type, respectively. Six patients had propionic academia with a prevalence of 1:310 200, 7 had isovaleric academia (1:265 900), 6 had glutaric academia type 1 (1:310 200), 27 had 3-methylcrotonyl-CoA carboxylase deficiency (MCC, 1:68 900), 1 had 3-hydroxy-3-methylglutaric aciduria (1:1 861 300), 2 had β-ketothiolase deficiency (1:960 600), and 3 had biotinidase deficiency/holocarboxylase synthetase deficiency (1:620 400). Thirty-one patients had a disease onset at neonatal period, and 15 at post-neonatal period. Thirty-three patients had brain involvements or cranial imaging disorders. Three patients with MMA had kidney diseases or heomlytic uremic syndrome, and 3 had myocardial impairments. Twenty patients died during the follow-up.</p><p><b>CONCLUSIONS</b>MMA is the most common newborn organic aciduria in Zhejiang province. Except MCC, most organic aciduria may lead to metabolism decompensation, complications or even death.</p>
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<p><b>OBJECTIVE</b>To screen fatty acid oxidation disorders (FAOD) in newborns in Zhejiang province.</p><p><b>METHODS</b>A total of 1 861 262 newborns were screened for FAOD in Neonatal Screening Center of Zhejiang Province during January 2009 and December 2016. The blood samples from newborns were screened by tandem mass spectrometry, and diagnosis of FAOD was confirmed by urine organic acid measurement combined with genetic analysis. The prognosis and follow-up of patients with FAOD were also evaluated.</p><p><b>RESULTS</b>Of 1 861 262 newborns screened, 121 cases of FAOD were diagnosed. Among 121 cases of FAOD, primary carnitine deficiency (PCD) was the most common type (=78, 64.5%), which was followed by short-chain acyl-CoA dehydrogenase deficiency (SCADD,=27, 22.3%), medium-chain acyl-CoA dehydrogenase deficiency (=5, 4.1%), multiple acyl-CoA dehydrogenase deficiency (MADD,=4, 3.3%), very long-chain acyl-CoA dehydrogenase deficiency (=3, 2.5%), carnitine palmitoyltransferase Ⅰ deficiency(=2, 1.7%)and carnitine palmitoyltransferase Ⅱ deficiency (CPTⅡD,=2, 1.7%). During 2-82 month follow-up, 15 patients were lost, 4 were dead (1 PCD, 1 MADD, and 2 CPTⅡD), and the remaining 102 subjects had normal intelligence and physical development without any clinical symptoms.</p><p><b>CONCLUSIONS</b>PCD and SCADD are the most common FAODs in newborns in Zhejiang province. Most of FAOD patients are asymptomatic, and have normal growth and development after early intervention and management.</p>
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Objective The aim of this study was to evaluate the value of serum exosomal miRNAs,originating from tumor tissue cells,could be used as noninvasive biomarker for distinguishing clear cell renal cell carcinoma (ccRCC).Methods 30 pairs of tissue samples and the corresponding serum samples were collected from 20 ccRCC male patients and 10 female ccRCC patients,operated in our department from June 2015 to June 2016.Their age ranged from 45 to 70 years old,mean 57 years old.Based on the miRNA microarray analysis of ccRCCs miRNA expression profiles,we picked up four miRNAs,including miR-210,miR-224,miR-452,and miR-34a,to confirm our hypothesis.Then the expression quantity of these four miRNAs in tissues,serums and serum exosomes were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).Sensitivity,specificity and area under curve (AUC) for serum miRNA levels were determined using receiver operator characteristic (ROC) analysis.Results Expression of miR-210,miR-224,and miR-452 were higher in tumor tissues compared to those in adjacent noncancerous tissues (P＜0.05),increasing by 20.51-fold,54.08-fold and 2.48-fold respectively.But only miR-210 was significantly higher in ccRCC patients compared to healthy controls (HCs) in serum and serum exosome (P ＜0.05),increasing by 2.45-fold and 2.32-fold respectively.ROC curve analysis indicated that the serum exosomal miR-210 level might serve as a useful biomarker for differentiating patients with ccRCC from those with HCs;the AUC was 0.8789 (95％ CI 0.7803-0.9775) and the sensitivity and specificity was 92.1％ and 80.0％,respectively.Conclusion The detection of miR-210 in the serum exosome is useful for early diagnosis of clear cell renal cell carcinoma.
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Objective To investigate the effects of physical and chemical factors in the environment for dried blood sample (DBS) preparation of neonatal screening assay.Methods A total of 60 normal and 120 positive DBS were prepared under control and 10 different conditions.Another 30 normal and 80 positive DBS were prepared under control and 7 different concentration gradients of formaldehyde.The levels of phenylalanine (Phe),glucose-6-phosphate dehydrogenease (G6PD),thyroid stimulating hormone (TSH) and 17α-hydoxyprogesterone (17α-OHP) were tested by time-resolved fluorescence immunoassay or fluorescence assay.Statistical analysis was performed using SPSS 22.0 software.Results Compared with the control group,the results of Phe were not significantly different (P ＞ 0.05) when the samples were dried under the formaldehyde sensitive threshold (4.62 to 6.95 ppm for 18 hours).G6PD levels were significantly lowered when the samples were dried under all the conditions except for fast cold drying (2 to 8 ℃ overnight and formaldehyde condition,0.30 to 0.38 ppm for 4 hours or 0.21 to 0.24 ppm for 18 hours).TSH and 17α-OHP levels were lowered obviously when the samples were dried under the conditions of humidity,UV and formaldehyde condition (TSH:0.32 to 0.52 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours,17α-OHP:4.37 to 4.62 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours).The results of Phe,G6PD,TSH and 17α-OHP were not statistically different with the control group when the samples were dried under the fast cold drying and 2 to 8 ℃ overnight.Conclusion The physical and chemical factors in the environment of DBS preparation should be related to the accuracy of neonatal disease screening closely.The necessary control factors including formaldehyde,ethanol,glacial acetic acid,ultraviolet irradiation,heat,humidity and decoration pollution may exhibit significant effects on the preparation of DBS.Fast cold drying and overnight at 2 to 8 ℃ could be available for DBS preparation.
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Objective The aim of this study was to evaluate the value of serum exosomal miRNAs,originating from tumor tissue cells,could be used as noninvasive biomarker for distinguishing clear cell renal cell carcinoma (ccRCC).Methods 30 pairs of tissue samples and the corresponding serum samples were collected from 20 ccRCC male patients and 10 female ccRCC patients,operated in our department from June 2015 to June 2016.Their age ranged from 45 to 70 years old,mean 57 years old.Based on the miRNA microarray analysis of ccRCCs miRNA expression profiles,we picked up four miRNAs,including miR-210,miR-224,miR-452,and miR-34a,to confirm our hypothesis.Then the expression quantity of these four miRNAs in tissues,serums and serum exosomes were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).Sensitivity,specificity and area under curve (AUC) for serum miRNA levels were determined using receiver operator characteristic (ROC) analysis.Results Expression of miR-210,miR-224,and miR-452 were higher in tumor tissues compared to those in adjacent noncancerous tissues (P＜0.05),increasing by 20.51-fold,54.08-fold and 2.48-fold respectively.But only miR-210 was significantly higher in ccRCC patients compared to healthy controls (HCs) in serum and serum exosome (P ＜0.05),increasing by 2.45-fold and 2.32-fold respectively.ROC curve analysis indicated that the serum exosomal miR-210 level might serve as a useful biomarker for differentiating patients with ccRCC from those with HCs;the AUC was 0.8789 (95％ CI 0.7803-0.9775) and the sensitivity and specificity was 92.1％ and 80.0％,respectively.Conclusion The detection of miR-210 in the serum exosome is useful for early diagnosis of clear cell renal cell carcinoma.
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Objective To study the effect of bacillus licheniformis and live combined bifidobacterium and lactobacillus on chemotherapy-induced diarrhea (CID). Methods Seventy-two gastric cancer patients with CID were collected and the clinical data were analyzed retrospectively. The patients were divided into 4 groups according to different treatment methods with 18 cases each. The patients in group A were treated with montmorillonite powder, the patients in group B were treated with montmorillonite powder and bacillus licheniformis, the patients in group C were treated with montmorillonite powder and live combined bifidobacterium and lactobacillus, and the patients in group D were treated with montmorillonite powder, bacillus licheniformis capsule and live combined bifidobacterium and lactobacillus. The patients with severe diarrhea in 4 groups were treated with rehydration, maintaining water and electrolyte balance and nutritional support. The Karnofsky performance scale score (KPS score), diarrhea grading before and after treatment and treatment effect were recorded. Results The KPS score after treatment in group A, group B, group C and group D were significantly higher than that before treatment: (70.6 ± 10.6) scores vs. (62.2 ± 12.2) scores, (76.1 ± 7.8) scores vs. (61.7 ± 9.9) scores, (77.2 ± 7.5) scores vs. (61.1 ± 10.8) scores, (83.9 ± 5.0) scores vs. (63.9 ± 10.9) scores. Moreover, The KPS score in group B, group C and group D were significantly higher than that in group A, the KPS score in group D was significantly higher than that in group B and group C, and there were statistical differences (P<0.05). The diarrhea grading after treatment in 4 groups were improved compared with that before treatment. Furthermore, the diarrhea grading in group B, group C and group D were significantly better than that in group B and group C, and there were statistical differences (P<0.05). In group A, there were 2 cases with excellent effect, 10 cases with effect and 6 cases with no effect;in group B, there were 8 cases with excellent effect, 8 cases with effect and 2 cases with no effect;in group C, there were 7 cases with excellent effect, 9 cases with effect and 2 cases with no effect; in group D, there were 9 cases with excellent effect, 9 cases with effect and 0 case with no effect. The treatment effect in group D was significantly better than that in the other 3 groups, and there were statistical differences (Hc = 10.81, P<0.05). Conclusions Bacillus licheniformis and live combined bifidobacterium and lactobacillus is more effective in the treatment of gastric cancer patients with CID.
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Newborn screening of inherited metabolic diseases by tandem mass spectrometry is flourishing in our country.With expansion of the coverage and spectrum of diseases, it is important to strengthen the quality management, optimize the performance and reduce the false rate currently, such as quality assurance of the specimens, quality control of the process, quality verification of the test procedure, quality evaluation of the laboratory and quality optimization of the interpretation method. Along with development of the equipment, software and project, promotion of regional collaboration by data and experience sharing will be more critical in the future, and national neonatal screening by tandem mass spectrometry will step into a new stage.
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OBJECTIVE@#To investigate the diagnosis applying effects of ocular vestibular evoked myogenic potentials(oVEMP) in peripheral BPPV disease.@*METHOD@#During September 2012 to January 2015, we selected 80 healthy people in our hospital medical center as the control group, choose the same period of primary benign paroxysmal positional vertigo as the observation group of 80 patients. Two groups were carried out fully functional auditory evoked potential analysis, determination of oVEMP and cervical vestibular evoked myogenic potentials (cVEMP) anomaly amplitude threshold, P1 latencies, N1 incubation period.@*RESULT@#The cVEMP abnormal rate in the observation group was 28.8%, the oVEMP abnormal rate was 38.8%, while cVEMP and oVEMP abnormal rates in the control group was 1.3% and 2.5% respectively that compared to significant differences between the two groups (P < 0.05). The oVEMP test amplitude in the observation group was (5.98 ± 2.15) µv, the N1 incubation period was (10.03 ± 0.76)ms, while the control group were (4.09 ± 2.11)µv and (11.67 ± 0.78) ms that compared difference were statistically significant (P < 0.05). The cVEMP test amplitude in the observation group was (154.8 ± 43.9)2 µv, while the control group was (180.49 ± 45.34)µv, compared the difference was statistically significant (P < 0.05).@*CONCLUSION@#Paroxysmal positional vertigo patients ocular vestibular evoked myogenic potentials abnormal rate is relatively high, the utricle dysfunction for more severe than the balloon can be the subject of an objective function of the ear stone judgment, judgment in favor of the disease.