BACKGROUNDVitrification is a prospective technology in ovarian tissue cryopreservation, but it is still in an initial stage. This study was conducted to investigate a modified vitrification protocol for human ovarian tissue, which can be used as an alternative to preserve fertility for young women with cancer who have to undergo cytotoxic therapy and sterilization.

METHODSOvarian tissue samples were collected from 15 patients and randomly allocated to groups of fresh, vitrification, and conventional slow freezing. A modified carrierless vitrification method was applied. The proportion of morphologically intact follicles in fresh ovarian tissues was compared with that in warmed/thawed tissues. The initial growth of the follicles and the concentrations of estradiol and progesterone were detected to determine the viability and endocrine function of the cryopreserved tissues.

RESULTSThe proportion of morphologically intact primordial follicles in the fresh group (97.6%) was significantly higher than that in the other two groups (vitrification group 80.3% and slow-freezing group 72.6%, P < 0.001). In both the vitrification and slow-freezing groups, estradiol and progesterone were secreted continuously during 2-week culture in vitro, the proportion of primary follicles were both significantly increased compared to the fresh group. No statistically significant differences existed between the two groups after cryopreservation in the proportion of both primordial and primary follicles, and the concentrations of estradiol and progesterone (P > 0.05).

CONCLUSIONThe modified vitrification method for cryopreservation of human ovarian tissues is effective, simple, and inexpensive.

Adult ; Cryopreservation ; methods ; Estradiol ; biosynthesis ; Female ; Humans ; Ovary ; cytology ; metabolism ; Progesterone ; biosynthesis ; Tissue Culture Techniques

OBJECTIVETo explore the genetic etiology of fetuses with ventricular septal defects (VSD) using chromosomal microarray analysis (CMA).

METHODSA total of 248 fetuses were divided into isolated VSD group, VSD with other cardiac and/or great vessels malformation group, VSD with extra-cardiac anomalies group (including malformation and sonographic soft markers), and VSD with both cardiac and extra-cardiac anomalies group. Standard karyotyping was carried out for all fetuses, and CMA was performed for 6 fetuses with an abnormal karyotype and a proportion of fetuses with a normal karyotype. All cases were followed up, and neonates were followed up until 1 year of age.

RESULTSChromosomal abnormalities were identified in 60 (24.2%) of the 248 fetuses. For 6 of the fetuses subjected to further CMA analysis, the origin of abnormal chromosomes were clarified, among which 2 have overlapped with the critical region of Wolf-Hirschhorn syndrome. Candidate genes for VSD included WHSC1, LBX1, LDB3 and BBS10. For 143 fetuses with a normal karyotype, CMA has identified pathogenic copy number variations (CNVs) in 11 cases (7.7%). These included 9 well-known microdeletion or microduplication syndromes, including 22q11.2 microdeletion, 17p11.2 microdeletion (Smith-Magenis syndrome), 17p13.3 microdeletion (Miller-Dieker syndrome), 1p36 microdeletion, 1q21.1 microduplication and 4q deletion. Candidate genes for VSD included TBX1, LZTR1, FAT1, AKAP10, SKI, PRDM26, GJA5, ERCC4 and YWHAE. For 48.7% of the fetuses with benign CNVs, spontaneously closure has occurred within the first year of life.

CONCLUSIONCMA may increase the detection rate of submicroscopic imbalances by 7.7%. No significant correlation between different groups of VSD and the pathogenic CNVs was observed. Whole-genome CMA should be recommended to the fetuses with VSD but a normal karyotype. Nearly half of VSDs with benign CNVs may close spontaneously within the first year of life.

Chromosome Aberrations ; Chromosome Deletion ; DNA Copy Number Variations ; Heart Septal Defects, Ventricular ; genetics ; Humans ; Infant ; Infant, Newborn ; Karyotyping ; Microarray Analysis ; methods ; Prenatal Diagnosis ; methods

OBJECTIVETo analyze the distribution of the human leukocyte antigen (HLA)-A, B and DRB1 allele and haplotype in cord blood samples preserved in Guangzhou Cord Blood Bank collected in the last 10 years.

METHODSThe HLA-A, B and DRB1 genotyping of 4194 cord blood samples were detected by Special Monoclonal Tray, PCR-sequence specific promer (PCR-SSP), PCR-sequence specific oligonucleotide probe (PCR-SSO) and sequence based typing (SBT). Frequencies of HLA-A, B and DRB1 allele and haplotype were calculated by Arlequin software.

RESULTSThe total numbers of HLA-A, B and DRB1 alleles are 18, 43, 13 respectively. The obviously high frequency alleles are A*11, A*02, A*24, A*33, B*40, B*15, B*46, B*13, DRB1*12, DRB1*15, DRB1*09 and DRB1*04, with accumulative frequency of each locus being more than 50%. The most common haplotypes are A2-B46, B46-DR9, A11-DR12 and A2-B46-DR9.

CONCLUSIONThe distribution of HLA-A, B and DRB1 allele and haplotype of cord blood in Guangzhou Cord Blood Bank has typical characteristics of southern Chinese Han population. Authors' data may help in searching for appropriate donors.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; Haplotypes ; Humans

OBJECTIVETo study the feasibility of using Narcotrend (NCT) in monitoring the anesthetic depth during endotracheal intubation in sevoflurane anesthesia.

METHODSThirty ASA I-II patients (aged 20-49 years) undergoing gynecologic surgery under general anesthesia with tracheal intubation were randomized into sevoflurane group (n=15) and sevoflurane plus rocuronium group (n=15). In the former group, anesthesia was induced with sevoflurane at the primary concentration of 8% till the final end expiratory concentration reaching 2 MAC(minimum alveolar concentration) for 3 min, followed then by tracheal intubation and further observation of the indicators for another 3 min. The patients in sevoflurane plus rocuronium group received identical anesthesia procedures except for the administration of intravenous injection of rocuronium (0.6 mg/kg) after the loss of eyelash reflex. The NCT, BIS and hemodynamics were recorded during the process.

RESULTSNo significant differences were noted in NCT, bispectral index (BIS), MAP and heart rate before tracheal intubation between the two groups (P>0.05). The NCT and BIS increased significantly after tracheal intubation in sevoflurane group (P<0.05), but remained below 60. No significant changes in NCT and BIS occurred during intubation in sevoflurane plus rocuronium group (P>0.05). The mean arterial pressure (MAP) and heart rate were significantly increased in both groups after tracheal intubation in comparison with those before tracheal intubation (P<0.05), but the increment in sevoflurane group was significantly greater (P<0.05).

CONCLUSIONNCT may reflect the changes of the anesthetic depth resulting from the nociceptive stimulus of tracheal intubation in sevoflurane- induced anesthesia. NCT and BIS can not serve such a purpose in combined anesthesia with sevoflurane and rocuronium.

Adult ; Androstanols ; administration & dosage ; Anesthesia ; Anesthetics, Intravenous ; administration & dosage ; Hemodynamics ; Humans ; Intubation, Intratracheal ; methods ; Methyl Ethers ; administration & dosage ; Middle Aged ; Monitoring, Intraoperative ; methods ; Young Adult