Prenatal diagnosis and genetic analysis of a fetus with partial deletion of Yq and mosaicism of 45,X / 中华医学遗传学杂志

OBJECTIVE@#To carry out prenatal diagnosis and genetic analysis for a fetus with disorders of sex development (DSDs).@*METHODS@#A fetus with DSDs who was identified at the Shenzhen People's Hospital in September 2021 was selected as the study subject. Combined molecular genetic techniques including quantitative fluorescence PCR (QF-PCR), multiplex ligation-dependent probe amplification (MLPA), chromosomal microarray analysis (CMA), quantitative real-time PCR (qPCR), as well as cytogenetic techniques such as karyotyping analysis and fluorescence in situ hybridization (FISH) were applied. Ultrasonography was used to observe the phenotype of sex development.@*RESULTS@#Molecular genetic testing suggested that the fetus had mosaicism of Yq11.222qter deletion and X monosomy. Combined with the result of cytogenetic testing, its karyotype was determined as mos 45,X[34]/46,X,del(Y)(q11.222)[61]/47,X,del(Y)(q11.222),del(Y)(q11.222)[5]. Ultrasound examination suggested hypospadia, which was confirmed after elective abortion. Combined the results of genetic testing and phenotypic analysis, the fetus was ultimately diagnosed with DSDs.@*CONCLUSION@#This study has applied a variety of genetic techniques and ultrasonography to diagnose a fetus with DSDs with a complex karyotype.

Analysis for 6-methyladenine modification of DNA in chorionic tissue from aborted fetuses with monosomy 21 / 中华医学遗传学杂志

OBJECTIVE@#To study the correlation of genome-wide distribution of 6-methyladenine (6mA) of DNA in chorionic tissues from abortuses with monosomy 21.@*METHODS@#Genomic DNA was extracted from chorionic samples from four abortuses with monosomy 21 and four without. After quality and purity test, partial DNA was subjected to chromatin immunoprecipitation with anti-6mA antibody, and then identified by sequencing. The sequencing data was analyzed by using bioinformatic software for the difference in 6mA between the two groups.@*RESULTS@#Analysis of read peaks suggested that the control group have much more 6mA genes (n=4607) compared with the experiment group (n=1059). For chromosome 21, this difference is even more pronounced (8032 vs. 1769). Above results suggested that the level of 6mA modification in monosomy 21 is low. Gene ontology enrichment analysis and KEGG pathway enrichment analysis indicated that the absence of 6mA genes in monosomy 21 is closely related to the growth and development of embryo.@*CONCLUSION@#The 6mA modification of human genes may play a similar role to 5-methylcytosine (5mC) modification during the growth and development of embryos.

Karyotyping analysis on umbilical vein cord blood lymphocytes in middle-late pregnant fetus / 中国医师杂志

Objective To investigate the significances of karyotyping analysis on umbilical cord vein blood lymphocytes in the diagnosis of abnormal karyotypes in middle to late period of pregnant fetus.Methods A volume (0.5 ～ 1 ml) of umbilical cord vein blood was extracted from pregnant women in third trimester pregnancy with prenatal detection indications,and collected in sterilized anticoagulant tube.Lymphocytes were cultured and collected for karyotyping analysis after fixed and dropped on slides.Data were analyzed statistically.Results Lymphocytes were cultured successfully in 1 211 cases out of total 1 213 cases collected.Totally 142 abnormal karyotypes were found,which includes 81 cases (detection rate 6.68 ％) of non-heteromorphic abnormal chromosomes and 61 cases (detection rate 5.03％) of heteromorphic chromosomes.Among these abnormal karyotypes,50 cases (accounting for 35.21％ in total abnormal cases) of aneuploidy include 4 cases of chimerical karyotype.Structural abnormalities were found in 31 cases (accounting for 21.83％ in total abnormal cases) samples including 11 cases of translocations,17 cases of inversion and 3 cases of deletion.Conclusions Based on our findings,karyotyping analysis on umbilical cord vein blood lymphocytes could be an effective method for detect abnormal karyotypes in middle to late period of pregnant fetus and played an important role in prenatal diagnosis.

The neuroprotective effect of electrical stimulation in chronic epilepsy / 中华物理医学与康复杂志

Objective To study any protection against hippocampal neuron damage induced by epilepsy (SE) provided by transcutaneous stimulation (TNS) of the trigeminal nerve and to document any effect of such stimulation on the expression of glutamic acid decarboxylase (GAD) 65/67.Methods Pilocarpine injection was used to induce epilepsy in healthy male Sprague-Dawley rats which were then randomly divided into a treatment group and a model group.Rats which had not received the pilocarpine injection served as normal controls.In the treatment group the rats were given electrostimulation for one month after the first spontaneous seizure following the injection of pilocarpine.In the model group they were given sham TNS for one month.After the month of stimula-tion,immunohistochemistry was used to detect the expression of GAD65/67 in the hippocampus.Terminal deoxy-nucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays and Nissl staining were applied to deter-mine apoptosis and neuron loss in the hippocampus.Results Significantly less apoptosis was observed in the treatment group than in model group at 24 h,48 h and 72 h post-injection.Compared to the model group,average GAD65/67 expression had increased significantly in the treatment group at 24 h,72 h,1 week,2 weeks and 4 weeks post-stimulation.GAD65 expression reached its peak from 72 h to 1 week post-stimulation,then decreased to the level of the control group by 4 weeks post-stimulation.The expression of GAD67 remained elevated at all the time points employed.Conclusions TNS can significantly protect hippocampal neurons from damage in epilepsy,at least in rats.The underlying anti-epileptic and neuroprotective mechanisms may involve increased inhibitory transmission induced by the stimulation.

Hyperbaric oxygen for suppressing glial scar formation and inflammation after a stab wound to the cerebral cortex / 中华物理医学与康复杂志

Objective To observe any influence of hyperbaric oxygen (HBO) treatment on the formation of glial scars,and to explore how HBO suppresses the inflammatory reaction to injury.Methods A total of 96 healthy,adult,male,Sprague-Dawley rats were used to model cerebral puncture injury.They were then randomized into a control group and a treatment group,with 48 rats in each group.The treatment group received HBO treatment,while the control group received no special treatment.At 1,3,7,14 and 28 days after the puncture injury,the rats' right brain tissues were harvested and immunohistochemical staining was employed to compare the changes in number of astrocytes and microglial cells around the injury in the two groups.The level of tumor necrosis factor α (TNF-α) and interleukin 1 β (IL-1β) in the cerebral tissue was examined using ELISA.Results Among the control group the average wound areas after 7,14 and 28 days were (2.73 ± 0.05)μm2,(3.42 ± 0.18)μm2 and (2.41 ± 0.09) μm2,a significant reduction after 28 days compared with 7 and 14 days.The corresponding average wound areas of rats in the treatment group were (2.78±0.12)μm2,(2.59 ±0.08)μm2 and (1.20 ±0.06)μm2.There the average wound area had decreased significantly after 14 days,and the further reduction after 28 days was also significant.The numbers of GFAP-positive astrocytes at 14 and 28 days had increased significantly compared with after 7 days in both the control group and the treatment group.The average number of GFAP-positive astrocytes in the control group at 28 days had decreased significantly compared with after 14 days.Compared with the control group at the same time points,the number of GFAP-positivc astrocytes in the treatment group was significantly less.After modeling,the number of ionized calcium-binding adapter molecule Ⅰ (Ibal)-positive microglial cells increased significantly,but there was a significant decrease in both the control and treatment groups by 7 days.The average number of Ibal-positive microglial cells in the treatment group was significantly less than in the control group at all of the time points.Compared with the first day after modeling,the TNF-α concentration of the controls at 3 and 7 days was significantly higher,but by the 7th day it was significantly lower than it had been after 3 days.The average IL-1β concentration in the control group and TNF-α concentration in the treatment group had increased by day 3,but then decreased by day 7.The IL-1β concentration of the treatment group declined gradually.The average TNF-α and IL-1 β concentrations of the treatment group were significantly lower than those of the control group at all of the time points.Conclusion HBO treatment has a relatively good curative effect on cerebral puncture injury.It can accelerate wound healing and reduce the formation of glial scars.Its mechanism could be related to the deactivation of astrocytes and microglia cells and reducing the levels of cell factors that promote inflammation.