Objective:To carry out cytogenetic and molecular genetic analysis for two infertile patients carrying rare small supernumerary marker chromosomes (sSMC).Methods:Two infertile patients who received reproductive and genetic counseling at CITIC Xiangya Reproductive and Genetic Hospital on October 31, 2018 and May 10, 2021, respectively were selected as the study subjects. The origin of sSMCs was determined by conventional G banding, fluorescence in situ hybridization (FISH) and copy number variation sequencing (CNV-seq). Microdissection combined with high-throughput whole genome sequencing (MicroSeq) was carried out to determine the fragment size and genomic information of their sSMCs. Results:For patient 1, G-banded karyotyping and FISH revealed that he has a karyotype of mos47, XY, del(16)(p10p12), + mar[65]/46, XY, del(16)(p10p12)[6]/48, XY, del(16)(p10p12), + 2mar[3].ish mar(Tel 16p-, Tel 16q-, CEP 16-, WCP 16+ ). CNV analysis has yielded a result of arr[GRCh37]16p12.1p11.2(24999364_33597595)×1[0.25]. MicroSeq revealed that his sSMC has contained the region of chromosome 16 between 24979733 and 34023115 (GRCh37). For patient 2, karyotyping and reverse FISH revealed that she has a karyotype of mos 47, XX, + mar[37]/46, XX[23].rev ish CEN5, and CNV analysis has yielded a result of seq[GRCh37]dup(5)(p12q11.2)chr5: g(45120001_56000000)dup[0.8]. MicroSeq results revealed that her sSMC has contained the region of chromosome 5 between 45132364 and 55967870(GRCh37). After genetic counseling, both couples had opted in vitro fertilization (IVF) treatment and preimplantation genetic testing (PGT). Conclusion:For individuals harboring sSMCs, it is vital to delineate the origin and structural characteristics of the sSMCs for their genetic counseling and reproductive guidance. Preimplantation genetic testing after microdissection combined with high-throughput whole genome sequencing (MicroSeq-PGT) can provide an alternative treatment for carrier couples with a high genetic risk.

Objective:To explore the application effect of cardiac respiratory arrest scenario simulation exercise combined with four-step disk method in the teaching practice of standardized nurses in the department of anesthesiology, and to provide teaching plan and reference for the training of core competence of standardized nurses.Methods:A randomized controlled study was conducted to select 127 standardized trained nurses in rotation anesthesia and intoxication department of West China Hospital, Sichuan University from July 2020 to December 2023 by cluster sampling method. A total of 64 first-year standardized training nurses from three batches of rotation anesthesiology department from July 2020 to December 2021 were selected as the control group, and the conventional teaching method was applied to their teaching practice. A total of 63 first-year standardized training nurses from three batches of rotation anesthesiology department in 2022 and from July to December 2023 were selected as the observation group. On the basis of routine teaching, the cardio-respiratory arrest scenario simulation exercise combined with four-step review method was implemented to carry out teaching practice. One week before leaving the course, the theoretical test scores, skill operation scores (6 operations including anesthesia machine, intravenous induration needle infusion, open sputum aspiration, defibrillation apparatus, simple respiratory sac ventilation and single cardiopulmonary resuscitation), post competency scores and teaching satisfaction scores of the two groups of standardized training nurses were assessed and compared.Results:The control group included 11 males and 53 females, aged (23.25 ± 0.78) years. In the observation group, there were 12 males and 51 females, aged (23.29 ± 0.81) years. The average score of skill operation in the observation group was (93.54 ± 0.92) points, which was higher than that in the control group (91.02 ± 1.08) points, and the difference between the two groups was statistically significant ( t=-14.10, P<0.01). The total theoretical score of the observation group (81.32 ± 3.84) points was higher than that of the control group (61.86 ± 2.05) points, and the difference between the two groups was statistically significant ( t=-35.69, P<0.01). The scores of self-evaluation of post competence and other evaluation and teaching satisfaction of the observation group were (95.81 ± 2.69), (94.22 ± 2.57) and (96.81 ± 2.75) points, which were higher than those of the control group (88.47 ± 5.56), (91.64 ± 3.10) and (91.52 ± 3.50) points. The differences between the two groups were statistically significant ( t=-9.49, -5.10, -9.47, all P<0.01). Conclusions:The application of heart-respiratory arrest scenario simulation and GRAI repetition method in the teaching practice of standardized nurses can improve the theoretical mastery level of standardized nurses and their operational performance, enhance their post competency, improve their teaching satisfaction, enhance their self-confidence and self-awakening ability, improve their learning enthusiasm, and their theoretical connection with practice level. To promote the improvement of teaching program and teaching quality, it is worth applying and popularizing in teaching practice.

Objective:To explore the application effect of cardiac respiratory arrest scenario simulation exercise combined with four-step disk method in the teaching practice of standardized nurses in the department of anesthesiology, and to provide teaching plan and reference for the training of core competence of standardized nurses.Methods:A randomized controlled study was conducted to select 127 standardized trained nurses in rotation anesthesia and intoxication department of West China Hospital, Sichuan University from July 2020 to December 2023 by cluster sampling method. A total of 64 first-year standardized training nurses from three batches of rotation anesthesiology department from July 2020 to December 2021 were selected as the control group, and the conventional teaching method was applied to their teaching practice. A total of 63 first-year standardized training nurses from three batches of rotation anesthesiology department in 2022 and from July to December 2023 were selected as the observation group. On the basis of routine teaching, the cardio-respiratory arrest scenario simulation exercise combined with four-step review method was implemented to carry out teaching practice. One week before leaving the course, the theoretical test scores, skill operation scores (6 operations including anesthesia machine, intravenous induration needle infusion, open sputum aspiration, defibrillation apparatus, simple respiratory sac ventilation and single cardiopulmonary resuscitation), post competency scores and teaching satisfaction scores of the two groups of standardized training nurses were assessed and compared.Results:The control group included 11 males and 53 females, aged (23.25 ± 0.78) years. In the observation group, there were 12 males and 51 females, aged (23.29 ± 0.81) years. The average score of skill operation in the observation group was (93.54 ± 0.92) points, which was higher than that in the control group (91.02 ± 1.08) points, and the difference between the two groups was statistically significant ( t=-14.10, P<0.01). The total theoretical score of the observation group (81.32 ± 3.84) points was higher than that of the control group (61.86 ± 2.05) points, and the difference between the two groups was statistically significant ( t=-35.69, P<0.01). The scores of self-evaluation of post competence and other evaluation and teaching satisfaction of the observation group were (95.81 ± 2.69), (94.22 ± 2.57) and (96.81 ± 2.75) points, which were higher than those of the control group (88.47 ± 5.56), (91.64 ± 3.10) and (91.52 ± 3.50) points. The differences between the two groups were statistically significant ( t=-9.49, -5.10, -9.47, all P<0.01). Conclusions:The application of heart-respiratory arrest scenario simulation and GRAI repetition method in the teaching practice of standardized nurses can improve the theoretical mastery level of standardized nurses and their operational performance, enhance their post competency, improve their teaching satisfaction, enhance their self-confidence and self-awakening ability, improve their learning enthusiasm, and their theoretical connection with practice level. To promote the improvement of teaching program and teaching quality, it is worth applying and popularizing in teaching practice.

Primary ciliary dyskinesia (PCD) is a congenital, motile ciliopathy with pleiotropic symptoms. Although nearly 50 causative genes have been identified, they only account for approximately 70% of definitive PCD cases. Dynein axonemal heavy chain 10 (DNAH10) encodes a subunit of the inner arm dynein heavy chain in motile cilia and sperm flagella. Based on the common axoneme structure of motile cilia and sperm flagella, DNAH10 variants are likely to cause PCD. Using exome sequencing, we identified a novel DNAH10 homozygous variant (c.589C > T, p.R197W) in a patient with PCD from a consanguineous family. The patient manifested sinusitis, bronchiectasis, situs inversus, and asthenoteratozoospermia. Immunostaining analysis showed the absence of DNAH10 and DNALI1 in the respiratory cilia, and transmission electron microscopy revealed strikingly disordered axoneme 9+2 architecture and inner dynein arm defects in the respiratory cilia and sperm flagella. Subsequently, animal models of Dnah10-knockin mice harboring missense variants and Dnah10-knockout mice recapitulated the phenotypes of PCD, including chronic respiratory infection, male infertility, and hydrocephalus. To the best of our knowledge, this study is the first to report DNAH10 deficiency related to PCD in human and mouse models, which suggests that DNAH10 recessive mutation is causative of PCD.
Humans ; Male ; Animals ; Mice ; Semen/metabolism* ; Dyneins/metabolism* ; Cilia/metabolism* ; Mutation ; Ciliary Motility Disorders/genetics*

OBJECTIVE: To summarize the genetic characteristics of 671 Chinese pedigrees affected with Duchenne/Becker muscular dystrophy (DMD/BMD). METHODS: Clinical data of the pedigrees were collected. Multiplex PCR, multiple ligation dependent probe amplification (MLPA), next generation sequencing (NGS), Sanger sequencing and long read sequencing were used to detect the variant of DMD gene in the probands and their mothers, and prenatal diagnosis was provided for high risk pregnant women. RESULTS: Among 178 pedigrees analyzed by multiplex PCR, 44 variants of the DMD gene were detected, with the genetic diagnosis attained in 110 pedigrees. Among 493 pedigrees analyzed by MLPA in combination with NGS or Sanger sequencing, 294 pathogenic/possible pathogenic variants were identified, among which 45 were unreported previously, and the genetic diagnosis attained in 484 pedigrees. Structural variants of the DMD gene were identified in two pedigrees by long-read sequencing. Among 444 probands, 341 have inherited the DMD gene variant from their mothers (76.8%). Among 390 women with a high-risk, 339 have opted to have natural pregnancy and 51 chose preimplantation genetic testing for monogenetic disease (PGT-M). The detection rate of neonatal patients and carriers following natural pregnancy was significantly higher than that for PGT-M. CONCLUSION Combined application of MLPA, NGS, Sanger sequencing and long-read sequencing is an effective strategy to detect DMD/BMD. PGT-M can effectively reduce the risk of fetuses. Above finding has expanded the spectrum of DMD gene variants and provided a basis for reproductive intervention for pregnancies with a high risk for DMD/BMD.
China ; Dystrophin/genetics* ; Exons ; Female ; Genetic Testing ; Humans ; Infant, Newborn ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne/genetics* ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

OBJECTIVE: To analyze the (CGG)n repeats of FMR1 gene among patients with unexplained mental retardation. METHODS: For 201 patients with unexplained mental retardation, the (CGG)n repeats of the FMR1 gene were analyzed by PCR and FragilEase RESULTS: For the 201 patients with unexplained mental retardation, 15 were identified with full mutations of the FMR1 gene. The prevalence of fragile X syndrome (FXS) in patients with unexplained mental retardation was determined as 7.5% (15/201). Prenatal diagnosis was provided for 6 pregnant women with pre- or full mutations. Analysis revealed that women with mental retardation and full FMR1 mutations exhibited a skewed XCI pattern with primary expression of the X chromosome carrying the mutant allele. CONCLUSION FXS has a high incidence among patients with unexplained mental retardation. Analysis of FMR1 gene (CGG)n repeats in patients with unexplained mental retardation can facilitate genetic counseling and prenatal diagnosis for their families. FMR1 gene (CGG)n repeats screening should be recommended for patients with unexplained mental retardation.
Female ; Fragile X Mental Retardation Protein/genetics* ; Fragile X Syndrome/genetics* ; Humans ; Intellectual Disability/genetics* ; Mutation ; Pregnancy ; Prenatal Diagnosis

AIM: To establish cell line FL-POL? - and to study the role of POL?(polymerase kappa) on genetic stability. METHODS: A mammalian expression vector expressing antisense POL? gene fragment pMAMneo -amp -- POL? was constructed by cloning the 1 690-1 918 fragment of POL? gene into the mammalian expression vector pMAMneo-amp - in antisense orientation. FL cells were fransfected with this antisense RNA expressing vector and selected by G418. Based on the shuttle-plasmid pZ189, the mutation assay was made. RESULTS: The spontaneous mutation frequency of supF tRNA gene in the plasmid replicated in the FL-POL? - was 11.2?10 -4 , while it was 4.9?10 -4 and 3.7?10 -4 in the control cells FL and FL-M , respectively. CONCLUSION: POL? playes an important role in maintenance of genetic stability.