Abstract A method based on dispersive solid phase extraction-high performance liquid chromatography-tandem mass spectrometry for determination of 19 kinds of nonprotein nitrogen compounds including melamine, cyromazine, amidinourea, aminotriazine, 3-aminotriazole, 4-aminotriazole, allantoin, cyanuric acid, dicyandiamide, thiourea, semicarbazide, L-leucine, L-isoleucine, L-arginine, L-hydroxyproline, L-theanine, ammeline, ammelide and guanidine in powdered formulas was presented. The nonprotein nitrogen compounds were degreased by chloroform and extracted by acetonitrile, with MgSO4 to remove water and C18 to clean up. The samples were separated on Merck ZIC HILIC column (150 mm í2. 1 mm, 5 μm, 20 nm) with gradient elution. The electrospray ionization was operated in the positive mode and negative mode, and monitored by the multiple reaction monitoring ( MRM) mode. Allation was quantified by external standard method and the other 18 kinds of nonprotein nitrogen compounds were quantified by internal standard method. All of the correlation coefficients (r) were higher than 0. 99. The limits of quantitation (LOQ) were 0. 05-5. 0 mg/kg, the average recoveries were between 82 . 2% and 115 . 0%, and the relative standard deviations were less than 20%.

Objective To analyze the rate of blood culture contamination,pollution path,the proportion of contaminated bacteria of First People′s Hospital of Kunming city from 2013 to 2015,and provide a basis for effective prevention and control of pollution.Methods A total of 34 713 cases of blood culture samples from 2013 to 2015,were retrospectively analyzed.Results A total of 2860 culture positive samples from 34 713 cases were found,from which 361 cases were polluted(1.04%).According to the classification of the year,the lowest rate of blood culture contamination was 2015 According to the classification in the quarter,there was no significant difference between the 4 seasons;According to the classification of internal and surgical systems,,the pollution rate of internal medicine system was 1.06%,while the surgical system was 0.96%.According to the classification of departments:the hemodialysis center has the highest pollution rate(2.71%),followed by ICU (2.23%).Galactophore Department has the highest pollution rate (2.30%) in the surgical system and followed by orthopedics(1.92%).According to the statistics of contaminated bacteria,Staphylococcus aureus was the highest,accounting for 32.41%,followed by Staphylococcus epidermidis,accounting for 31.30%.Conclusion Hospital has a high blood culture contamination rateand diversification pollution waywhich can not be ignored.The pollution of bacteria in the blood culture mainly for bottle of single positive coagulase negative Staphylococcus.

A strain, Escherichia coli AC-521, which could efficiently covert glycerol to lactic acid, was isolated from soil samples. A series of morphological and biochemical characteristics and sequence analysis of 16S rDNA reveal that it belongs to Escherichia coli. Through orthogonal experiment of L16(45), the optimum medium compositions were determined as initial glycerol 70 g/L, yeast extract 4 g/L, peptone 7 g/L, (NH4)2SO4 10 g/L, and K2HPO4 2.5 g/L. The 5 L fermenter fed-batch fermentation was performed under optimal conditions, giving 74.5 g/L lactic acid with a yield of 0.87 mol/mol glycerol after 80 hours.

OBJECTIVE: To analyze the prenatal diagnosis procedure for a 45,X male fetus. METHODS: A 31-year-old women underwent amniocentesis due to a moderate risk of trisomy 21. The fetal cells were subjected to chromosomal karyotyping, BACs-on-Beads (BoBs) assay, chromosomal microarray analysis and fluorescence in situ hybridization. RESULTS: Combined analyses revealed that the whole of Yp has translocated to 21p, which yielded a fetal karyotype of 45,X,dic(Y;21)(q11;p11).ishdic(Y;21)(SRY+,CEPY+;CEP21+). CONCLUSION BoBs and modified N-banding method are helpful for the diagnosis of 45,X male fetus with Yp translocation.

OBJECTIVE: To report on a case of mosaicism 13q inversion duplication, analyze its mechanism, and discuss the correlation between its genotype and phenotype. METHODS: Amniotic fluid and umbilical cord blood were collected at 23 and 32 weeks of gestation, respectively. Combined with G-banding chromosome karyotyping analysis, single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) were used to confirm the result. RESULTS: The karyotype of the fetus was determined as 47,XY,+inv dup(13)(q14.3q34)/46,XY. After careful counseling, the couple decided to continue with the pregnancy, and had given birth to a boy at 40 weeks' gestation. Except for a red plaque (hemangioma) on the nose bridge, no obvious abnormality (intelligence to be evaluated) was discovered. CONCLUSION To provide reference for clinical genetic counseling and risk assessment, the location and proportion of new centromere formation should be fully considered in the case of mosaicism 13q inversion duplication.
Amniocentesis ; Chromosome Inversion/genetics* ; Comparative Genomic Hybridization ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Mosaicism ; Pregnancy ; Prenatal Diagnosis

Objective:To calibrate the absorbed doses of the configured ray water with different gears of energies in accelerator based onof International Atomic Energy Agency(IAEA)277 and 381 reports,so as to ensure the accuracy of the output dose of the linear accelerator during clinical radiotherapy.Methods:Elekta Infinity linear accelerator was used in this study,and the photon beam energies were respectively 6MV flattening filter(FF)mode and 6MV flattening filter free(FFF)mode.The electron beam energies were respectively 4,6,8,10,12 and 15MeV.According to the IAEA TRS277 and TRS381 reports,the calibrations of output doses in photon and electron beam waters were performed respectively by using the PTW dosimeter,PTW30013 finger type of ionization chamber and PTW34001 parallel plate ionization chamber.The error of each step was analyzed,and the accuracies of the calibrations of using different standards for the output waters of linear accelerator were compared.Results:The output amounts of photon beams of FF mode and FFF mode of 6MV at the maximum dose point in water were respectively 1.003 and 1.008cGy/MU.The output amounts of the energy of each gear of electron beams of 4,6,8,10,12 and 15MeV at the maximum dose point in water were respectively 1.003,1.002,0.998,0.999,1.000 and 1.003 cGy/MU.The calibration of the output of each gear of energy rays at the maximum dose point in water was 1cCy corresponded to 1MU,which error was less than 1%.Conclusion:The calibration for the output dose amount of accelerator in water on the basis of IAEA TRS277 and trs381 can ensure the accuracy of the output dose of the accelerator.

OBJECTIVE: To optimize the condition for chromosome flaking of mesenchymal stem cells to ensure the cytogenetic quality control of expanding production and clinical application. METHODS: Chromosomal flaking methods were optimized from current chromosome preparation techniques from the aspects of MSCs cell culture concentration, colchicine treatment time and low permeability time. RESULTS: By repeated pre-experiments, the optimal MSCS chromosome flaking condition of MSCs was determined as cell culture concentration of (1-2)× 10 cells per T25 cell culture bottle, and the colchicines processing time was determined as 2 hours and 10 minutes, and the low permeability was 1 hour. CONCLUSION The optimized chromosome flaking condition can fulfill the requirement of cytogenetic quality control for MSCs.
Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Chromosome Disorders ; Chromosomes, Human ; Cytogenetics ; Humans ; Mesenchymal Stem Cells

OBJECTIVETo explore the genetic etiology of three families affected with split-hand/split-foot malformation (SHFM).

METHODSPeripheral venous blood samples from 21 members of pedigree 1, 2 members of pedigree 2, and 2 members of pedigree 3 were collected. PCR-Sanger sequencing, microarray chip, fluorescence in situ hybridization (FISH), real-time PCR, and next-generation sequencing were employed to screen the mutations in the 3 families. The effect of the identified mutations on the finger (toe) abnormality were also explored.

RESULTSMicroarray and real-time PCR analysis has identified a duplication in all patients from pedigrees 1 and 3, which have spanned FKSG40, TLX1, LBX1, BTRC, POLL and FBXW4 (exons 6-9) and LBX1, BTRC, POLL and FBXW4 (exons 6-9) genes, respectively. A missense mutation of the TP63 gene, namely c.692A>G (p.Tyr231Cys), was found in two patients from pedigree 2. FISH analysis of chromosome 10 showed that the rearrangement could fita tandem duplication model. However, next-generation sequencing did not identify the breakpoint.

CONCLUSIONThe genetic etiology for three families affected with SHFM have been identified, which has provideda basis for genetic counseling and guidance for reproduction.

Chromosomes, Human, Pair 10 ; genetics ; Female ; Foot Deformities, Congenital ; genetics ; Genetic Testing ; Hand Deformities, Congenital ; genetics ; Humans ; Limb Deformities, Congenital ; genetics ; Male ; Mutation ; genetics ; Pedigree

Objective To investigate the clinical features of cutaneous adverse reactions to tyrosine kinase inhibitors.Methods Thirty patients with cutaneous adverse reactions to tyrosine kinase inhibitors were enrolled from the First Affiliated Hospital of Guangzhou Medical University between January 2015 and December 2016,and their laboratory test results,histopathological findings and treatment response data were collected and analyzed retrospectively.Results Of the 30 patients,15 presented with acneiform eruptions,10 with eczematoid eruptions,2 with morbilliform rashes,1 with telangiectasia,1 with hand-foot skin reaction,9 with xerosis,7 with nail changes and 4 with hair changes.A patient with grade 4 acneiform eruptions showed a markedly elevated alanine transaminase (ALT) level (315 U/L).Mild ALT abnormalities (48.5-88.1 U/L) were found in 3 patients with grade 3 acneiform eruptions,1 with grade 2 acneiform eruptions,1 with grade 1 acneiform eruptions and 1 with eczematoid eruptions complicated by fever.Two patients with eczematoid eruptions and 1 with morbilliform rashes showed elevated proportions of peripheral blood eosinophils (0.057-0.303).Pathological changes of the acneiform eruptions included hyperkeratosis and dilation of hair follicles and neutrophilic infiltration.Pathological manifestations of eczematoid eruptions included different degrees of spongiosis,thickened spinous layer,irregular elongation of rete ridges and liquefaction degeneration of basal cells in the epidermis,and perivascular infiltration of lymphocytes and eosinophils in the superficial dermis.Patients with grade 1-3 acneiform eruptions received oral minocycline for 6 weeks,skin lesions gradually regressed,but relapse occurred after the withdrawal.After withdrawal of targeted antineoplastic agents and 2-week treatment with systemic glucocorticoids,skin lesions gradually regressed in patients with grade 4 acneiform eruptions,those with eczematoid eruptions complicated by fever,and those with morbilliform rashes.Skin rashes also resolved in patients with mild morbilliform rashes and those with mild eczematoid eruptions after 2 weeks of treatment with antianaphylactic agents and topical glucocorticoids.Oral antibiotics were effective for the treatment of periungual erythematous swelling or granulomas.Conclusion Tyrosine kinase inhibitor-related cutaneous adverse reactions include a constellation of disorders,and hepatic function can be impaired.

OBJECTIVE: To present on a prenatally diagnosed case with complex structural rearrangements of chromosome 8. METHODS: Chromosome karyotyping, chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH) were carried out for a fetus with increased nuchal thickness. RESULTS: The karyotype of the amniotic fluid sample showed extra materials on 8p. FISH revealed a centromeric signal at the terminal of 8p with absence of telomeric signal. CMA revealed partial deletion of 8p23.3 [(208049_2256732)×1], partial duplication of 8p23.3p23.2 [(2259519_3016818)×3], and partial duplication of 8q [8q11.1q12.2(45951900_60989083)×3]. CONCLUSION The complex structural rearrangements of chromosome 8 in this case has differed from the commonly seen inv dup del(8p).
Female ; Pregnancy ; Humans ; Chromosomes, Human, Pair 8/genetics* ; In Situ Hybridization, Fluorescence ; Gene Rearrangement ; Prenatal Diagnosis ; Centromere