Genetic analysis of a case with mosaicism complex structural aberration of chromosome 18 / 中华医学遗传学杂志

OBJECTIVE@#To determine the karyotype of a patient with mosaicism complex structural aberration of chromosome 18.@*METHODS@#A male patient with a 2-year history of infertility presented at the Center of Reproductive Medicine of the Third Hospital of Peking University in October 2019 was selected as the study subject. Clinical data of the patient was collected. Peripheral blood sample was taken for chromosomal karyotyping, copy number variation (CNV) analysis and fluorescence in situ hybridization (FISH) assay. Semen sample was taken for single sperm CNV analysis.@*RESULTS@#The patient was found to have a karyotype of mos 47,XY,del(18)(q21q23),+r(18)(q21q23)[84]/46,XY,del(18)(q21q23)[9]/48,XY,del(18)(q21q23),+r(18)(q21q23)×2[6]/47,XY,del(18)(q21q23),+r(18)(q21q23×2)[1].ish 47,XY,del(18)(q21q23),+r(18)(q21q23)[84]/46,XY,del(18)(q21q23)[9]/48,XY,del(18)(q21q23),+r(18)(q21q23)×2[6]/47,XY,del(18)(q21q23),+r(18)(q21q23×2)[1]del(18)(q21q23)(D18Z1+,18p+,18q+,WCP18+),r(18)(q21q23)(WCP18+),r(18)(q21q23×2)(WCP18+). No pathogenic CNV was identified. Sequencing of 20 single sperms showed that 1 sperm was normal, 1 had yielded no result, 9 had harbored del(18q), 7 had harbored dup(18q)×2, and 2 had harbored dup(18q)×3. The dup/del fragments had both spanned approximately 33 Mb.@*CONCLUSION@#It is rare for carriers of complex structural and numerical abnormalities of chromosome 18 to have a normal phenotype. Based on the accurate cytogenetic and molecular analyses and the single sperm CNV analysis, the influence of the aberrant karyotype on the gametogenesis may be evaluated.

The construction of an organized hospital scientific research platform under the background of organized scientific research in hospital / 中华医学科研管理杂志

Objective:To explore the paths and goals of organized construction for scientific research platforms in large-scale hospitals under the background of organized scientific research in China.Methods:By reviewing the relevant literature, and analyzing the construction of existing international and domestic research platforms for organized scientific research, this article elaborated on the importance and necessity of building a research platform for clinical hospitals under the background of organized scientific research and made suggestions for the platform construction.Results:Organized scientific research requires the organized construction of scientific research platforms. The construction of organized scientific research platforms should always focus on the major national needs, serve major scientific plans, carry out organized talent training, and internal efficient and orderly organization and operation, under the principle of interdisciplinary and multi-level collaborative innovation and development.Conclusions:In response to the strategic needs of national science and technology in the 14-Five Year Plan, the hospital scientific research platforms should be upgraded, integrated, expanded, and improved in an organized way, to form a multi-disciplinary and cross-dimensional platform structure to ensure the smooth development of organized scientific research.

Expert consensus on preimplantation genetic testing for spinal muscular atrophy / 中华医学遗传学杂志

Spinal muscular atrophy (SMA) is an autosomal recessive motor neuron degenerative disease, which is the most common fatal neuromuscular disease in pediatrics with a high carrier frequency and can lead to progressive symmetrical muscle weakness and atrophy of the trunk and limbs. Preimplantation genetic testing (PGT) can be used to prevent the birth of children with SMA effectively. To standardize PGT technologies for SMA, experts from the fields of neurology, pediatrics and reproductive genetics have discussed and drafted this consensus for guiding its clinical application.

Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase II oocytes / 医学前沿

Oocyte cryopreservation is widely used for clinical and social reasons. Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures, but not storage time, can alter the gene expression profiles of frozen oocytes. Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase II oocytes remain unknown. Four women (30-32 years old) who had undergone IVF treatment were recruited for this study. RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes (3, 3, 4, and 3 oocytes were cryostored for 1,2, 3, and 12 months) were analyzed at a single-cell resolution. A total of 1987 genes were differentially expressed in the 13 vitrified-thawed oocytes. However, no differentially expressed genes were found between any two groups among the 1-, 2-, 3-, and 12-month storage groups. Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development. Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself, suggesting that long-term cryostorage of human oocytes is safe.

Advance in the methods of preimplantation genetic diagnosis for single gene diseases / 中华医学遗传学杂志

More than 7000 single gene diseases have been identified and most of them lack effective treatment. As an early form of prenatal diagnosis, preimplantation genetic diagnosis (PGD) is a combination of in vitro fertilization and genetic diagnosis. PGD has been applied in clinics for more than 20 years to avoid the transmission of genetic defects through analysis of embryos at early stages of development. In this paper, a review for the recent advances in PGD for single gene diseases is provided.

Effect of vitrification at the germinal vesicle stage on the global methylation status in mouse oocytes subsequently matured in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is still unclear whether the vitrification procedure itself is associated with the incidence of abnormal DNA methylation during oocytes vitrification. The purpose of this study was to evaluate the epigenetic profile of mouse oocytes, which went through vitrification either at a mature stage or at an immature stage following in vitro maturation (IVM) by analyzing the global DNA methylation.</p><p><b>METHODS</b>Metaphase II (M II) stage and germinal vesicle (GV) stage oocytes were collected from adult female mice and were vitrified respectively. The M II oocytes were assessed for cryo-survival and global DNA methylation. The GV oocytes were assessed for cryo-survival and only the surviving GV oocytes were cultured in vitro for subsequent assessment of global DNA methylation in mature oocytes. In vivo matured fresh M II oocytes without undergoing vitrification were used as control. The level of global DNA methylation in the M II oocytes was then examined by immunofluorescence using an anti-5-methylcytosine (anti-5-MeC) monoclonal antibody and fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG under a laser scanning confocal microscope.</p><p><b>RESULTS</b>In terms of the effect of vitrification on global DNA methylation status in matured oocytes, in the M II-v group, all the examined oocytes (90/90) were found with hypermethylation, including 63.3% (57/90) of them displaying DNA methylation of a very high level, 25.6% (23/90) with a high level, and 11.1% (10/90) with an intermediate level, whereas in the GV-v group, all the matured oocytes (129/129) were also examined with hypermethylation, including 67.4% (87/129) of them displaying DNA methylation of a very high level, 23.3% (30/129) with a high level, and 9.3% (12/129) with an intermediate level. Statistically, it was similar between both groups, which were similar to the control: 68.6% (83/121) of fresh M II oocytes displayed DNA methylation of a very high level, 21.5% (26/121) with a high level, and 9.9%(12/121) with an intermediate level (P > 0.05). In terms of the effect of IVM on global DNA methylation status in matured oocytes, in the in vivo matured oocytes group, all oocytes examined (94/94) were found with hypermethylation, including 80.9% (76/94) displaying DNA methylation of a very high level and 19.1% (18/94) with a high level, whereas in the in vitro matured oocytes group, all oocytes examined (69/69) were also found with hypermethylation: 85.2% (56/69) of them displayed with DNA methylation of very high level, 11.9% (11/69) with high level, and 2% (2/69) with intermediate level. This result was similar to that in in vivo matured fresh M II oocytes (P > 0.05).</p><p><b>CONCLUSION</b>The vitrification procedure at GV stage does not induce widespread alteration of global DNA methylation status of mouse oocytes subsequently matured in vitro.</p>

Application of the xenogenic acellular dermal matrix membrane application used in the postoperative tissue shortage repair / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To observe the short-term and long-term curative effect of the xenogenic acellular dermal matrix membrane (or joint muscle flap transfer) application used in the 82 cases postoperative tissue shortage repair that after the head neck carcinoma resection.@*METHOD@#To held the 82 cases head neck carcinoma postoperative mucosa shortage repaired after resection by the xenogenic acellular dermal matrix membrane (or joint muscle flap transfer), 65 cases mucosa shortage wound be directly covered by the repair membrane and the other 17 cases mucosa shortage wound be repaired by the tranfered muscle tissue flap with the repair membrane covered; 53 cases underwent additional postoperative radiotherapy between 2-4 weeks and follow-up in 1, 3, 6, 12, 18, 24, 30, 36, 48, 60 months and observed the operation site repair process through the electronic laryngoscope, observed the patients respiration, swallow, phonation function.@*RESULT@#Seventy-seven cases patients operation incision reached I phase healing standard, another 5 cases patients operation incision reached II phase healing standard because of the wound infection and fully-recovered through the local wound drainage,dressing process. All the patients tracheal cannula,the stomach tube be extubated successfully and without the local cicatricial constriction occurred. Seventy-eight cases follow up period reached 1 year including 53 cases who underwent postoperative radiotherapy, 49 cases follow up period reached 3 years including 32 cases who underwent postoperative radiotherapy, 14 cases follow up period reached 5 years including 12 cases who underwent postoperative radiotherapy. The patients with static local lesions discovered no reaction such as exclusion, allergy.@*CONCLUSION@#The application of xenogenic acellular dermal matrix membrane (or joint muscle flap transfer used in in the postoperative tissue shortage repair that after the head neck carcinoma resection have several advantage such as comparatively easily implementation, operation safety edge enough,well preserved organ function, comparatively low incidence about the laryngeal stenosis, the short-term and long-term repair effect are all exact.

Endoscope thyroid lobectomy through subclavicular approach / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the clinical applications of endoscopic thyroid lobectomy through subclavicular approach and to summarize the surgical experiences.@*METHOD@#Thirty-five cases were subjected to endoscopic thyroid lobectomy through subclavicular approach. Briefly, operative cavities were established by automatic retractors and the affected lobes were dissected and removed using suction dissectors combined with the harmonic scalpels. The inferior thyroid artery was used as the mark to locate the recurrent laryngeal nerve.@*RESULT@#All the operations achieved successfulness with no complication. 33 out of 35 were intraoperatively diagnosed by frozen-pathology as goiter and the last two cases were diagnosed as papillary thyroid carcinoma. There were no postoperative swelling, numbness or other kinds of uncomfortableness with benign patients and they were satisfied with the cosmetic outcomes.@*CONCLUSION@#Endoscopic thyroid lobectomy through subclavicular approach for thyroid benign lesions has the advantages of minimal invasion, high safety, easy manipulation and good cosmetic outcomes, which are worth of wide applications.

Inhibitory effect of silencing hTERT gene on growth of human squamous cell carcinoma xenograft in nude mice / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#This study is to explore the inhibitory effect of silencing hTERT gene by short-hairpin RNA on growth of nasopharyngeal carcinoma xenograft in nude mice with RNAi technique.@*METHOD@#Construction and expression of hTERT cDNA sequence according to the specific hTERT mRNA, including fluorescein eukaryotic expression vector, packaged into a lentivirus. qPCR and Western blot analyzed hTER.T mRNA and protein levels in transfected cells. Proliferation rate of transfected cells was determined by MTT assay in vitro. Cell growth cycle was detected by flow cytometry. The invasiveness of each group was compared using in vitro cell invasion assay.@*RESULT@#RT-PCR and Western blot analysis showed that, hTERT siRNA significantly reduced hTERT mRNA and protein levels, especially hTERT siRNA1. siRNA treatment inhibited tumor cell proliferation, and cell migration and invasiveness were significantly lower. Tumor cell growth rate was significantly different between control group and siRNA group (P 0.05).@*CONCLUSION@#Lentivirus containing specific sequences of hTERT gene could significantly inhibit the growth of nasopharyngeal carcinoma cells line. hTERT siRNA expression vector can effectively inhibit NPC cell proliferation, migration and invasion, which may provide a novel molecular targets for gene therapy of nasopharyngeal carcinoma.

Study in safety of total thyroidectomy / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the complications in total thyroidectomies and the safety of total thyroidectomy.@*METHOD@#Retrospective analyses 351 cases underwent total thyroidectomy in our department. Preoperative and postoperative electronic laryngoscopy were used to assess the vocal cord function. All patients have serum calcium analysis on the operative day and 3rd and 7th day after operation.@*RESULT@#Temporary hypocalcaemia occurred in 47.01% after total thyroidectomy, but only 15.67% with severe symptom temporarily,and 21.37% with temporary mild numbness. Permanent hypoparathyroidism occurred in 0.85% patients. None with bilateral recurrent laryngeal nerve palsy, and temporary unilateral vocal cord palsy occurred in 1.42% patients and permanent unilateral recurrent laryngeal nerve palsy occurred in 0.56% patients.@*CONCLUSION@#Total thyroidectomy is safe with low complications.

Mechanism of Hair Cell Apoptosis in Age-related Hearing Loss Mouse Cochlea / 听力学及言语疾病杂志

Objective In this study,we investigated the hair cell apoptosis and the molecular mechanisms in age-related hearing IOSS mouse cochlea.Methods All animals were assessed by ABR and DPOAE analysis in both ears.TUNEL(Terminal dexynucleotidyl transferase(TdT)mediated dUTP Nick End Labeling)and PI were used to identify DNA fiagments and caspase-3 activities in hair cells.Results The results showed that the nmf308 mice had progressive hair cell loss along with age.The cochlear OHCs were reduced 5%～10% at 1 month and 100% at 3 month in the basal region.Substantial amounts of TUNEL-postive OHCs nuclei were observed at age of 1 month,and activated caspase-3 labeling OHCs were most obviously observed at age of 2 months.Conclusion These results suggest that DNA single strand break is attributed primarily to apoptosis of cochlear lesion,and activation of caspase-3 at leter stage leads to the hair cell apoptiovs of nuclear condensation in age-related hearing loss mouse cochlea.

Clinical analysis of 43 cases with carcinomas in maxillary sinus by sequent combination therapy / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the clinical effects and life quality of 43 cases with carcinomas in maxillary sinus by sequent combination therapy.@*METHOD@#All the patients were subjected sequent and combination therapy, which included inductive chemotherapy through superficial temporal artery, operation and radical radiation therapy, and of these, 18 patients were added consolidation chemotherapy after combination therapy. Three year survival rate, 5- year survival rate and life quality were observed in a 5 years follow up.@*RESULT@#All patients completed this therapy, the overall 5-year survival rates were 46.5% (20/43) and 3-year were 65.1% (28/43). None were undertaken orbital exenteration and 12 patients recovered their ability of work and manual labour.@*CONCLUSION@#The sequent combination therapy is an effective method to enhance the prospective efficacy of treating the carcinoma of maxillary sinus and can raise the local controlling rate. The induced chemotherapy through the temporal artery superficial has many advantages, such as high density of topical remedy, obvious tumor regression after only a cycle chemotherapy, not delaying the surgery opportunity and so on. The consolidation chemotherapy after this combined therapy can reduce the risks of distant metastasis further.

The value of exposure recurrent laryngeal nerve in thyroidectomy / 中国耳鼻咽喉头颈外科

OBJECTIVE To explore the advantage of exposure recurrent laryngeal nerve in thyroidectomy. METHODS Exposure the recurrent laryngeal nerve was performed deliberately in 452 cases during thyroidectomy. The larynx was examined with laryngoscope before and after operation. RESULTS Of the 452 cases with 748 exposed recurrent laryngeal nerve, only 14 cases (1.88%) occurred temporary recurrent laryngeal nerve palsy, and 2 cases (0.27%) occurred permanent recurrent laryngeal nerve palsy. CONCLUSION Exposure the recurrent laryngeal nerve during thyroidectomy can protect the nerve from surgery injury.