Fetal skeletal dysplasia is a diverse group of degenerative diseases of bone and cartilage disorders that can lead to movement disorder and even death. This study aims to evaluate the diagnostic yield of sonographic examination and genetic testing for fetal skeletal dysplasia. From September 2015 to April 2021, the study investigated 24 cases with suspected short-limb fetuses, which were obtained from Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology. To identify the causative gene, multiple approaches (including karyotype analysis, copy number variations and whole exome sequencing) were performed on these fetuses. And further segregation analysis of the candidate variant was performed in parents by using Sanger sequencing. ① Out of 24 cases, likely pathogenic variants in FGFR3, FBN2, COL1A2, CUL7 and DYNC2H1 were detected in 6 cases; pathogenic variants in FGFR3, IMPAD1 and GORAB were identified in other 6 cases; and variants in WNT1, FBN1, OBSL1, COL1A1, DYNC2H1 and NEK1, known as Variant of Undetermined Significance, were found in 4 cases. There were no variants detected in the rest 8 cases by the whole exome sequencing. ② Of 24 cases, 12 (50

Background: It is challenging to make an accurate prenatal diagnosis for congenital anomalies of the kidney and urinary tract (CAKUT) because of its pathologic diversity. This study aims to evaluate the performance of whole-exome sequencing (WES) combined with karyotype analysis and copy number variations (CNVs) in diagnosing high-risk fetal CAKUT.Methods: We conducted a retrospective study on prenatal diagnoses of CAKUT in our hospital from January 2020 to April 2021. The research studied 24 high-risk fetuses with CAKUT who were scanned by ultrasonography at the prenatal diagnosis center of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology. The likely pathogenic gene variants were screened for the patients and their parents by multiple approaches, including karyotype analysis, CNVs and WES, and further verified with Sanger sequencing.Results: ①We detected abnormal CNVs in 20.8% (5/24) of the fetuses but only 8.3% (2/24) fetuses had abnormal karyotypes. ②Of the 15 CAKUT fetuses, positive findings (40%) were detected by WES. Of the 9 high-risk fetuses with CAKUT (negative findings in ultrasound scan but with family history), we found abnormal variants (77.8%) through WES.Conclusion: The application of CNVs and WES showed advance in prenatal diagnosis of CAKUT and the pathogenic gene variants were detectable especially for high-risk fetuses with negative ultrasound findings on CAKUT in the preliminary study. The applied strategy could be used to improve the accuracy of prenatal diagnosis for CAKUT in the future.

A novel heterozygous mutation (c.325dup) was identified in EXT1 gene from the proband and the affected family members; this mutation was absent in all the unaffected family members. The identification of the novel frameshift insertion mutation (c.325dup) expands the mutation spectrum of HME, which provides new evidence for HME diagnosis.

To identify the pathogenic gene variation in a Chinese family with Hereditary Multiple Exostoses (HME). By examining blood-sourced DNA and clinical manifestations of the proband and his family members, the whole exome sequencing (WES) and Sanger sequencing were used to detect possibly pathogenic mutations. A novel heterozygous mutation (c.325dup) was identified in exon 1 of the exostosin 1 (EXT1) gene from the proband and the affected family members. And we found this mutation was absent in all the unaffected family members. This c.325dup mutation is in the exon 1 domain of the EXT1 gene and the change of p.C109Lfs*80 cause the early termination of protein translation. The identification of the novel frameshift insertion mutation (c.325dup) expands the mutation spectrum of HME, which provides new evidence for HME diagnosis.

目的:利用全外显子测序(WES)技术分析胎儿肠管增宽的致病性突变,为产前诊断及遗传咨询提供帮助.方法:对3例肠管增宽的胎儿进行染色体核型分析、拷贝数变异(CNV)及WES检测.结果:3例胎儿染色体核型分析结果均正常,1例胎儿CNV检出15号染色体微重复,3例胎儿WES结果均发现与肠管增宽相关的基因突变.病例1为GDNF基因c.329G>A杂合变异,遗传自其父亲,可引起先天性巨结肠症3型,该病为常染色体显性遗传,并具有低外显率的特点;病例2为NOTCH2基因的c.1310A>C新发变异,与Alagille综合征2型、Hajdu-Cheney综合征相关,均为常染色体显性遗传;病例3为SLC26A3基因复合杂合突变,包括来自父亲的c.1427del突变和来自母亲的c.269_270 dup突变,可导致常染色体隐性遗传的先天性失氯性腹泻1型.经Sanger验证病例1为临床意义未明突变,根据既往病史判断该突变可能致病,病例2及病例3均为致病性突变.结论:WES技术对于分析胎儿肠管增宽的遗传变异具有重要的诊断价值.

The proband is a five-year-old boy diagnosed with Duchenne muscular dystrophy (DMD) by clinical manifestations and laboratory examination, but clinical phenotype of his parents is normal. In the study, his mother had a second pregnancy, and they went to obstetrics for genetic counseling to make informed reproductive choices.

The Duchenne Muscular Dystrophy (DMD) gene variants are associated with the disease phenotypes. The pathogenic mutation, c.2293-1G>C, was detected in DMD gene in the proband and the fetus, which has not been reported in the literature.The minigene expression in vitro confirmed that c.2293-1G>C is responsible of aberrant splicing.

目的 探讨双绒毛膜双羊膜囊双胎妊娠一胎畸形在孕中晚期实施经腹选择性药物减胎术的临床特点、减胎时机、安全性及妊娠结局.方法 回顾性分析2017年5月至2019年3月华中科技大学同济医学院附属同济医院产科收治的12例双胎妊娠一胎畸形经腹选择性减胎孕妇的临床资料.结果 所有接受选择性减胎术的孕妇绒毛膜性均为双绒毛膜双羊膜囊,减胎时平均孕周为(24.68±3.22)周.受孕方式为自然受孕6例,体外受精-胚胎移植辅助生殖技术5例,使用促排卵药物1例.减胎指征:系统胎儿超声筛查提示一胎儿严重畸形者9例,产前诊断提示一胎儿染色体异常者5例.减胎方式均采取B超引导下经腹胎儿心脏或脐静脉内注射10％氯化钾溶液,减胎成功率为100％,未见感染、流产、死胎等并发症.妊娠结局:4例发生早产,8例足月分娩,新生儿Apgar评分均在7分以上.结论 对双绒毛膜双羊膜囊的双胎妊娠一胎畸形的孕妇于孕中晚期实施经腹选择性药物减胎术是安全可行的,但对操作人员的技术有较高要求,尽早实施减胎术有利于成功减除畸形胎儿、得到良好的妊娠结局,对家庭及社会都有重要意义.

目的:探讨胎儿肠管增宽的临床表现与妊娠结局之间的相关性,为临床咨询、判断预后和指导治疗提供依据.方法:回顾分析2015年10月至2019年6月于华中科技大学附属同济医院胎儿系统超声诊断为肠管增宽的51例孕妇的临床资料.结果:51例孕妇中,19例妊娠结局良好,32例出现不良妊娠结局(引产、胎死宫内、新生儿死亡、出生后需手术治疗等),占62.7％.胎儿十二指肠增宽的不良结局发生率为100％,明显高于小肠增宽(54.2％)及结直肠增宽(25.0％).孕32周前诊断胎儿肠管增宽不良妊娠结局发生率为72.7％,而孕32周后检出者为44.4％.胎儿肠管增宽合并羊水过多22例,不良结局发生率为86.4％,明显高于羊水量正常孕妇(40.0％).10例孕妇行羊膜腔穿刺产前诊断均未发现异常,2例十二指肠增宽新生儿行基因检查为21-三体综合征.结论:产前超声诊断胎儿肠管增宽与消化系统畸形及妊娠结局密切相关,及时发现、密切随访、早期处理极为重要.

目的:探讨肾移植术后妊娠患者的孕期管理及分娩方式.方法:回顾分析4例肾移植术后妊娠患者的妊娠过程、结局、分娩方式和围产儿结局.结果:4例女性患者均接受同种异体肾移植术,分别于术后5～16年成功妊娠.2例患者早孕期超声发现胚胎停止发育.2例患者孕期给予FK506(他克莫司)、美卓乐(甲泼尼松龙片)和依木兰(巯唑嘌呤片)治疗,FK506浓度2～9ng/ml,其中1例患者孕38+1周行剖宫产术分娩一活女婴,1例孕35+5周胎膜早破,早产经阴分娩一活女婴.随访1～2年,婴幼儿均健康.结论:肾移植术后患者本身及胎儿风险较高,应视为高危妊娠.妊娠前应更换合理的免疫抑制方案,孕期监测FK506浓度,精确调整免疫抑制剂及激素剂量,使其药物浓度保持在治疗窗内,确保患者顺利妊娠及分娩.

Liddle 综合征(Liddle Syndrome)又称假性醛固酮增多症,是一种罕见的单基因致病型高血压,遗传方式为常染色体显性遗传,遗传学基础是编码ENaC 的基因(SCNN1B 和SCNN1G)发生功能获得性突变,主要特征包括高血压、低钾血症、低肾素及低醛固酮血症等,常于青少年起病.该病常有家族聚集性,也存在散发情况,由 Grant Liddle 等于 1963年首次进行详细描述.现将我们收集的一个家系的临床特点、基因检测结果及产前诊断报告如下.

Mammary serine protease inhibitor (maspin; also known as serpin family B member 5 (SERPINB5)) plays a vital role in regulating the biological functions of extravillous trophoblast (EVT) cells, but the mechanism remains unclear. Vascular endothelial growth factor (VEGF) C is a signature angiogenic molecule expressed and secreted by first-trimester trophoblasts, and bioinformatics analyses has revealed upregulation of VEGFC in pre-eclampsia. The aim of this study was to explore whether maspin regulates EVT cells by regulating the expression of VEGFC. Reverse transcription-polymerase chain reaction and western blotting were used to investigate the effects of hypoxia on the expression of VEGFC in EVT cells. Cells were treated with recombinant (r) maspin and decitabine (to selectively inhibit DNA methyltransferases and then upregulate maspin gene expression), and the effects on VEGFC expression evaluated. In addition, the effects of rVEGFC on the biological functions of EVT cells invitro were evaluated using cell migration and invasion assays. Hypoxia increased the expression of VEGFC in EVT cells. rMaspin upregulated the expression of VEGFC in normoxic EVT cells, and downregulated the expression of VEGFC in hypoxic EVT cells at 24h. Decitabine increased VEGFC expression in normoxic EVT cells, but had no significant effect on VEGFC expression in hypoxic EVT cells. rVEGFC promoted the migration and invasion of normoxic EVT cells and inhibited the invasion of hypoxic EVT cells. These results suggest that VEGFC is involved in the regulation of maspin in EVT cell migration and invasion. However, other molecular mechanisms may be involved and require further investigation.

目的:利用高通量测序(NGS)技术分析家族性Alport综合征(AS)的致病性突变及遗传方式,为咨询者提供准确的遗传咨询并给予产前诊断.方法:对3例家族性AS先证者进行致病性突变分析及家系验证,对遗传咨询者进行产前诊断.结果:3例先证者均发现AS致病基因COL4 A5点突变,突变位点分别位于c2633 G→A、c1769 A→C、c1352G→A,经家系验证均为致病性突变.对3例咨询者胎儿DNA进行Sanger验证(第一代DNA测序技术),提示1例胎儿携带AS致病基因COL4 A5的错义突变,2例未检测出致病性突变.结论:认识AS遗传方式的多样性和遗传特征,强调重视先证者的基因诊断及家系验证,确定遗传方式,建议行遗传咨询并给予生育指导.

Objective:To study and summarize the experience of the treatment to acute aortic dissection with gestation.Methods:The clinical data of 4 pregnant patients with acute aortic dissection during May 2006 to Nov.2016 were retrospectively analyzed.One of cases received cesarean section and total aortic arch replacement and desending aortic stenting (Betall''s operation) under general anesthesia.One of cases received Bentall operation after she operated cesarean section three days.The other woman underwent abdominal aortic replacement after cesarean section two months,and the last case was treated with cesarean section and aortic dissection endovascular replacement.4 cases of patients retained uterus,and 3 cases of the newborn survived.Results:After 6 months of follow-up,the computerized tomographic angiography(CTA) examination of the patients with aortic dissection showed the formation of false lumen thrombosis(1 case was not recovered because she left hospital less than1 month).4 pregnant women were successfully recovered,3 cases of the survival newborn were normal,and 1 case of newborn died.Conclusion:CTA and echocardiography for aortic dissection pregnancy timely and accurate diagnosis is crucial.Intraoperative and postoperative should ensure maternal hemodynamics.According to the gestational age and aortic dissection type and other factors,we decides to terminate the timing and operative methods of pregnancy.

Objective: Pre-eclampsia is a major cause of maternal mortality and morbidity. Conditions with low oxygen tension are regarded as a key factor. Decitabine can partly attenuate the effects of hypoxia. This research was designed to investigate the effects of decitabine in rats with NG-Nitro-L-arginine Methyl Eater (L-NAME) induced pre-eclampsia and to explore the molecular mechanisms. Methods: A Wistar rat model of pre-eclampsia was established by intraperitoneal injection of L-NAME, and the intervention reagent was decitabine. Blood pressure (BP) and 24-h urinary protein were monitored. The expression of Mammary Serine Protease Inhibitor (SERPINB5, maspin) in the placenta was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting. Results: Systolic BP in the tail artery of pregnant rats was increased by more than 30mm Hg, and 24-h urinary protein was significantly increased after L-NAME was added. After decitabine treatment, blood pressure and 24-h urinary protein were significantly decreased. The expression of SERPINB5 in the placenta significantly increased after L-NAME was added. Decitabine significantly elevated the expression of SERPINB5 in the placenta of rats with L-NAME-induced preeclampsia. Conclusion: Decitabine reduced 24-h urinary protein and partly decreased blood pressure of pre-eclampsia in late pregnancy in rats with L-NAME-induced pre-eclampsia and increased the expression of SERPINB5, but the molecular mechanism of decitabine's effect remains unknown. This research provided a potential approach to studying the pathogenesis, treatment and prevention of pre-eclampsia.

Objective:To study the mechanism of low weight molecular heparin treating proteinuria of rats with L-NAME induced preeclampsia by synaptopodin. Methods:We dynami-cally monitored the blood pressure of tail artery,the total amount of 24h urinary protein and the mean body weight of pups of pregnancy rats after different concentration of low molecular weight heparin ( LMWH) administration. Meantime we detected synaptopodin expression of kidneys in pregnant rats after different concentration low molecular weight heparin ( LMWH ) administra-tion using RT-PCR, Immunohistochemical staining and Western blot. Results:Compared with the control,synaptopodin expression of kidneys in pregnant rats was significantly decreased after L-NAME injection. Synaptopodin expression of kidneys in pregnant rats was significantly in-creased after high concentration of LMWH ( H-LMWH ) administration compared with the L-NAME group. But synaptopodin expression had no changes between the L-NAME group and low concentration of LMWH ( L-LMWH) group. In addition,the blood pressure of tail artery and proteinuria of pregnant rats with L-NAME injection were markedly deceased after low and high concentration of LMWH administration [(200IU/(kg·d) and 600IU/(Kg·d)] on the 15th and 19th day. But the body weights of pups and the number of embryos per female did not change among the three groups. Conclusion:LMWH improved proteinuria of pregnant rats with L-NAME induced preeclampsia by up-regulation of synaptopodin,while LMWH improved pro-teinuria and decreased blood pressure of tail artery.

Objective Extravillous trophoblast (EVT) cells invade the endometrium and the maternal spiral arterioles during the first trimester. Mammary Serine Protease Inhibitor (Maspin, SERPINB5) plays a putative role in regulating the invasive activity of cytotrophoblasts. The maspin gene is silenced in various cancers by an epigenetic mechanism that involves aberrant cytosine methylation. We investigated the effect of the methylation status of the maspin promoter on the maspin expression and the aggressiveness of EVT cells. Methods Western blotting was used to detect the maspin protein expression in EVT cells upon hypoxia. The proliferative ability, the apoptosis rate and the migration and invasiveness were measured with Cell Counting Kit-8 assay, Flow Cytometry technology and Transwell methods. Subsequently, we treated cells with recombinant maspin protein. The methylation degree of maspin promoter region upon hypoxia/ decitabine was detected by bisulfite sequencing PCR and methylation-specific PCR. Finally, we explored the effects of decitabine on maspin protein expression and the aggressiveness of EVT cells. Results Hypoxia effectively increased maspin protein expression in EVT cells and significantly inhibited their aggressiveness. The addition of recombinant maspin protein inhibited this aggressiveness. Decitabine reduced the methylation in the maspin promoter region and effectively increased the maspin protein expression, which significantly weakened the migration and invasiveness of EVT cells. Discussion The methylation status of the maspin promoter is an important factor that affects the migration and invasion of EVT cells during early pregnancy. A decrease in the methylation status can inhibit the migration and invasion of EVT cells to affect placentation and can result in the ischemia and hypoxia of placenta.