Application of single sperm sequencing for the preimplantation genetic testing of a Chinese family affected with Spinal muscular atrophy / 中华医学遗传学杂志

OBJECTIVE@#To assess the value of single sperm sequencing in preimplantation genetic testing for monogenic disease (PGT-M).@*METHODS@#A Chinese couple with two children whom had died of Spinal muscular atrophy (SMA) and attended the Jiangxi Provincial Maternal and Child Health Care Hospital in June 2020 was selected as the subject. Eleven single sperm samples were isolated by mechanical immobilization and subjected to whole genome amplification. Real-time PCR and Sanger sequencing were used to detect the SMN1 variants in the single sperm samples. Genomic DNA of the wife, her parents and the husband, as well as one single sperm sample harboring the SMN1 variant and two single sperm samples without the variant were used for the linkage analysis. Targeted capture and high-throughput sequencing were carried out to test 100 single nucleotide polymorphisms distributed within 2 Mb up- and downstream the variant site. The haplotypes linked with the SMN1 variants were determined by linkage analysis. Blastocyst embryos were harvested after fertilizing by intracytoplasmic sperm injection. Cells from the trophoblasts of each embryo were biopsied and subjected to whole genome amplification and targeted capture and high-throughput sequencing to determine their carrier status. Chromosomal aneuploidy of wild-type embryos was excluded. An euploid embryo of high quality was transferred. Amniotic fluid sample was taken at 18 weeks of gestation to confirm the status of the fetus.@*RESULTS@#Genetic testing showed that the couple both had deletion of exons 7 ~ 8 of the SMN1 gene. The wife has inherited the deletion from her father, while the husband was de novo. The haplotypes of the husband were successfully constructed by single sperm sequencing. Preimplantation genetic testing has indicated that 5 embryos had harbored the heterozygous variant, 4 embryos were of the wild type, among which 3 were euploid. Prenatal diagnosis during the second trimester of pregnancy has confirmed that the fetus did not carry the deletion.@*CONCLUSION@#By single sperm sequencing and PGT-M, the birth of further affected child has been successfully avoided.

Effectiveness, safety and cost of urinary follicle stimulating hormone in controlled ovarian stimulation in China: multi-center retrospective cohort study of 102 061 in vitro fertilization cycles / 中华妇产科杂志

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

Expression and significance of osteopontin and muscle segment homeobox gene Msx2 in sinonasal inverted papilloma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression level and significance of osteopontin (OPN) and muscle segment homeobox2 (Msx2) in sinonasal inverted papilloma (SNIP), and the relationship with the process of SNIP malignant transfomation.@*METHOD@#Immunohistochemical method was used to detect the expression of osteopontin and Msx2 in 32 cases of sinonasal inverted papilloma (SNIP), 30 cases of inflammatory nasal polyp paraffin tissue (INP) and 30 cases of sinonasal inverted papilloma with carcinoma. According to the pathology results SNIP were divided into mild dysplasia, moderate dysplasia, severe dysplasia. All the datas were analyzed by SPSS13.0.@*RESULT@#The positive expression of OPN and Msx2 in the SNIP with carcinoma were all 100% (30/30), were significantly higher than the SNIP and INP, and the pairwise comparisons were all statistically significant (P < 0.05). The expression of OPN and Msx2 in the SNIP were divided according to pathological. There was significant difference between mild dysplasia and severe dysplasia group (P < 0.05), while there were not significant difference between light degree of dysplasia and moderate dysplasia group, moderate dysplasia and severe dysplasia group. And the expression of OPN and Msx2 was positively correlated in SNIP (P < 0.01).@*CONCLUSION@#OPN and Msx2 may play an important role in the pathway of progression of SNIP, which may be identified as the new therapeutic targets.

Application of autogenous bone pate for atticus reconstruction and mastoid cavity obliteration in tympanoplasty with mastoidectomy / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the curative effect of autogenous bone pate for atticus reconstitution and mastoid cavity obliteration in canal wall down mastoidectomy with tympanoplasty.@*METHOD@#The 60 patients (60 ears) of cholesteatomatous tympanitis were randomly divided into 2 groups. The experimental group underwent canal wall down mastoidectomy with tympanoplasty and using autogenous bone pate for atticus reconstitution and mastoid cavity obliteration. The control group only underwent canal wall down mastoidectomy with tympanoplasty. All patients were followed up for 0.5-2 years. Improvement of hearing and dry ear time in these two groups was retrospectively observed.@*RESULTS@#There are significant difference between two groups in improvement of hearing and dry ear time, according to follow-up and analysis of therapeutic efficacy (P < 0.05).@*CONCLUSION@#Hearing can be improved, dry ear time can be shorten and correlative complication can be decreased by atticus reconstruction and mastoid cavity obliteration with autogenous bone pate in canal wall down mastoidectomy with tympanoplasty, which guarantees the normal activity space of auditory bone chain.

Expression and significance of Msx2 and topo II-alpha in sinonasal inverted papilloma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expression and significance of muscle segment homeobox2 (Msx2) and topo II-alpha in sinonasal inverted papilloma (SNIP), and the relationship in the process of malignant transformation of SNIP.@*METHOD@#Immunohistochemical method was used to detect the expression of Msx2 and topo II-alpha in 32 cases of SNIP, 30 cases of inflammatory nasal polyp (INP) and 30 cases of SNIP with carcinoma. According to the pathology results, SNIP were divided into mild atypical hyperplasia, moderate atypical hyperplasia and severe atypical hyperplasia.@*RESULT@#The mean optical density of Msx2 in SNIP and SNIP with carcinoma tissues were 0.2183 +/- 0.0598 and 0.2521 +/- 0.0761,which were significantly higher than 0.1878 +/- 0. 0372 in the INP tissue (P<0.05 or 0.01). The mean optical density of topo II-alpha in SNIP and SNIP with carcinoma tissues were 0.2303 +/- 0.0397 and 0.2666 +/- 0.0483, which were significantly higher than 0.1978 +/- 0.0388 in the NIP tissue (P<0.01). There were significant difference of Msx2 and topo II-alpha in SNIP between any two of the three groups divided according to pathological morphology (P<0.01 or 0.05). The expression of Msx2 and topo II-alpha in SNIP were positively correlated (P<0.05).@*CONCLUSION@#Msx2 and topo II-alpha may play an important role in the occurrence and development of SNIP. So it can be used as new therapeutic targets.

Psychological Status of Patients with Gynecological Malignant Tumors and Analysis of Correlated Factors / 中国肿瘤临床

Objective: To investigate the psychosocial characteristics and mind-physique status of pa-tients with gynecological malignant tumors. Methods: Clinical psycho-rating scale [life event scale (LES) and symptom checklist-90 (SCL-90)] was applied to test life events and mind-physique status of 60 cases of pa-tients with gynecological malignant tumor. The results were compared with those of 40 female patients with benign tumors and 40 normal females. Results: The frequency and tension value of total life events and those of negative life events in the malignant cancer group were much higher than those in the benign tumor group and normal group (P<0.05) and no remarkable differences were found in the frequency of positive life events among the three groups. The frequencies of total life events and negative life events were higher in the be-nign tumor group than those in the normal group (P<0.05) but there were no significant differences in the fre-quency of positive life events, total tension value or positive tension value among the three groups. No signifi-cant differences were found in the indices of SCL-90, positive scores and scorns of other factors among the three groups (P>0.05). The scores of somatization, obsession, interpersonal relationship sensitivity, anxiety, hostility, phobia, and paranoia were higher in the malignant tumor group than in the benign tumor group and normal group (P<0.05). The average positive scores and the depression scores in the benign tumor group were higher than those in the normal group (P<0.05), but there was no statistical significance in other indices between the benign tumor group and the normal group (P>0.05). Conclusion: Negative psychosocial factors are closely correlated with the occurrence and growth of gynecological malignant tumors. Active psychological intervention should be performed as eady as possible.