A) in COL1A1 gene resulting in OI in a Chinese family. The detailed molecular and clinical features will be useful for extending the evidence for genetic and phenotypic heterogeneity in OI and exploring the phenotype-genotype correlations in OI.

Objective To report the prenatal molecular diagnosis for two gravida in a family with spondylepiphyseal dysplasia congenita(SEDC)caused by G504S mutation of COL2A1 gene.Methods DNA of the two fetuses was extracted from amniotic fluid at the 19+3 and 18+6 weeks of gestation respectively.Direct sequencing of two samples were performed after amplifying exon 23 of COL2A1 containing the potential mutation.The femur length and biparietal diameter of the first fetus were measured by sonographic scans every two weeks from 17+3 weeks to 27+3 weeks of gestation,and for the second fetus these parameters were measured from 16+1 to 19+1 weeks of gestation.Results Sequncing analysis revealed the first fetus and his mother presented the same mutation which is specifically associated with SEDC,but the second fetus did not show the mutation of COL2A1 gene.Biparietal diameters of the both fetuses were appropriate for gestational age.Femur length of the second fetus was normal for gestational age but that of the first fetus was shortened evidently after the 23 week of gestation.The parents of the first fetus determined to terminate the pregnancy.A medical termination was carried out at 27+5 weeks of gestation and a male fetus with a relatively large head and short limbs was delivered.The radiological findings of the fetus were consistent with SEDC including generalized platy spondesand shortened long bones.Conclusions Prenatal molecular diagnosis is important for the fetus with risk of SEDC and useful for genetic counseling.Genotype of fetus with risk of SEDC can be identified before sonographic scan.Molecular genetic analysis in conjunction with sonographic monitoring was helpful in prenatal diagnosis of SEDC.

Objective: To report a case of azoospermia with a karyotype of 45,X,der(Y)t(Y;13)(q11.2;q12),-13,accompanied with slight bilateral gynecomastia and multiple nodules.Methods: The karyotype was identified by karyotyping and FISH,and the breakpoints of the Y chromosome and the copy number of the BRCA2 gene in 13q12 determined by PCR-STS and DNA polymorphic analysis.The testis and nodule tissues of the patient were obtained for biopsy.Results: FISH confirmed SRY and centromere of the Y chromosome on the questionable 13 chromosome and the karyotype to be 45,X,der(Y)t(Y;13)(q11.1;q12),-13.ish der(Y)(SRY+,DYZ3+,wcp13+).PCR-STS showed the deletion of regions AZFa,b and C,with a breakpoint located inYq11.1 below sY82.No deletion of the BRCA2 gene was observed.The patient was diagnosed with Sertoli cell-only syndrome by testicular biopsy and with angiolipomata by pathological examination of the nodule tissue.Conclusion: The patient's phenotype of complete masculinization could be attributed to presence of the SRY gene,and his azoospermia with small testis to the absence of a fragment from Yq11.1 to Yqter.However,the molecular mechanism of angiolipoma remains unknown.

Objective To report a familial dentinogenesis imperfecta type Ⅱ (DGI type Ⅱ) with a novel splicing mutation in DSPP (dentin sialophosphoprotein) gene.Methods Based on the result of linkage analysis performed previously to map the candidate gene DSPP in the family, the promoter,the first four exons and exon-intron boundaries of DSPP were directly sequenced for the members of the DGI type Ⅱ family. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to confirm the results of sequencing.Results A novel splicing mutation of 23 bp deletion in intron 2 of DSPP gene was identified by DNA sequence analysis. The mutation changed acceptor site sequence from CAG to AAG, and might result in functional abolition of possible branch point site in intron 2. DHPLC result was consistent with that of sequencing. The mutation may be identified in all affected individuals, but not found in normal members of the family and 50 controls.Conclusion These results suggest the deleted mutation of DSPP gene causes DGI type Ⅱ in the family. The mutation has not been reported before.

Objective To examine the dosimetric advantages of three-dimensional (3D) computed tomography (CT)-guided interstitial brachytherapy (BT) for target volume and surrounding normal tissue in patients with locally advanced cervical cancer,and to provide a simple and effective clinical treatment approach.Methods A total of 52 patients who had poor tumor response to external beam radiotherapy (EBRT) with a residual tumor greater than 5 cm at the time of BT were included.The patients were treated by 3D CT-guided interstitial BT using a hybrid applicator comprised of uterine tandem and free metal needles.The high-risk clinical target volume (HR-CTV),intermediate-risk clinical target volume (IR-CTV),and organs at risk (OAR) were contoured.The total dose,including external beam radiotherapy and high dose-rate BT,was biologically normalized to conventional 2 Gy fractions (EQD2).D90and D100for both HR-CTV and IR-CTV,and D2 ccfor the bladder,rectum,and sigmoid were analyzed.Results The mean D90value for HR-CTV was 88.4±3.5 Gy.The D2 ccfor the bladder,rectum,and sigmoid were 81.1±5.6,65.7±5.1,and 63.1±5.4 Gy,respectively.D2 cc≤90 Gy for the bladder and D2 cc≤70 Gy for the sigmoid were observed in all the patients.D2 cc≤70 Gy for the rectum was observed in 89% of patients.Conclusions 3DCT-guided interstitial BT has a significant dosimetric advantage for target volume accompanied by few minor complications,and thereby may be clinically feasible for treating locally advanced cervical cancer.However,its long-term efficacy and possible toxicities will require further clinical observation.

OBJECTIVETo determine the origin of chromosomal aberration for a child featuring multiple malformation, and to correlate the genotype with phenotype.

METHODSRoutine G-banding was performed to analyze the karyotype of the patient and her parents, and array comparative genomic hybridization (array CGH) was used for fine mapping of the aberrant region.

RESULTSThe karyotype of the child was ascertained as 46,XY. Array CGH has mapped a 14.21 Mb deletion to 5p15.2p15.33, and a very small 3.67 Mb duplication to 5q35.3. The patient has presented features such as mental retardation, heart defect, low-set ears, hypertelorism and down-slanting palpebral fissures.

CONCLUSIONChromosome 5 copy number variation can cause multiple malformation. In contrast to routine karyotype analysis, array CGH can map aberrant region with much higher resolution and accuracy.

Abnormalities, Multiple ; diagnosis ; genetics ; Chromosome Aberrations ; Chromosomes, Human, Pair 5 ; DNA Copy Number Variations ; Genotype ; Humans ; Infant ; Male ; Phenotype

Objective To investigate the dosimetric difference between inverse planning simulated annealing(IPSA)and manual optimized plan for isodose line in interstitial brachytherapy for locally advanced cervical cancer and to provide a better optimization method for clinical application. Methods A total of 104 patients with cervical cancer were enrolled in this study. They received pelvic external beam radiotherapy and interstitial brachytherapy in five fractions. Both IPSA and manual optimized plan for isodose line were used to optimize the dose in each fraction. Dose volume parameters of the two plans were compared to analyze the dosimetric outcome by paired t-test. Results There were no significant differences in mean D 90and D 100for high-risk clinical target volume(HR-CTV)and D 90for intermediate-risk clinical target volume(IR-CTV)between the two groups(P>0.05). The IPSA group had a significantly higher D 100for IR-CTV than the manual optimized group(58.36±2.06 Gy vs. 53.99±2.17 Gy, P=0.025). For organs at risk,the IPSA group had a significantly lower mean rectum D 2ccand a significantly higher bladder D 2ccthan the manual optimized group(68.53± 2.85 Gy vs. 71.77± 1.79 Gy, P=0.002;80.49± 3.36 Gy vs. 78.71± 2.64 Gy,P=0.034). There was no significant difference in sigmoid D 2ccbetween the two groups(P>0.05). The IPSA group had significantly higher relative dose homogeneity index(HI)and conformity index (CI)of radiation dose for target volume than the manual optimized group(P<0.05), and there was no significant difference in overdose volume index(OI)between the two groups(P= 0. 1 0 7).Conclusions Compared with manual optimized plan for isodose line, IPSA can improve the dose distribution of tumor tissue,reduce mean rectum D 2cc,and increase CI and HI,so it is a preferable optimized treatment planning method in clinical application.

OBJECTIVE: To explore the genetic basis for a child with multiple malformations. METHODS: A child who had presented at Shanxi Provincial Children's Hospital in February 2021 was selected as the study subject. Clinical data of the patient was collected, and whole exome sequencing (WES) was carried out to screen pathogenic variants associated with the phenotype. Candidate variant was validated by Sanger sequencing of her family members. RESULTS: The child had normal skin, but right ear defect, hemivertebral deformity, ventricular septal defect, arterial duct and patent foramen ovale, and separation of collecting system of the left kidney. Cranial MRI showed irregular enlargement of bilateral ventricles and widening of the distance between the cerebral cortex and temporal meninges. Genetic testing revealed that she has harbored a heterozygous variant of NM_178014.4: c.217A>G (p.Met73Val) in the TUBB gene, which was unreported previously and predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). The child was diagnosed with Complex cortical dysplasia with other brain malformations 6 (CDCBM6). CONCLUSION CDCBM is a rare and serious disease with great genetic heterogeneity, and CDCBM6 caused by mutations of the TUBB gene is even rarer. Above finding has enriched the variant and phenotypic spectrum of the TUBB gene, and provided important reference for summarizing the genotype-phenotype correlation of the CDCBM6.
Humans ; Child ; Female ; Abnormalities, Multiple ; Blood Group Antigens ; Family ; Malformations of Cortical Development/genetics* ; Brain ; Mutation

【Objective】 To analyze the genetic variation characteristics and clinical phenotypes of a family with primary microcephaly (MCPH) caused by RTTN gene variation, and to provide reference for genetic counseling and prenatal diagnosis. 【Methods】 Clinical data of the three patients (including 2 fetuses and 2-year-old proband,and one fetus with clinical diagnosis) and their parents were collected and analyzed. Two of the children and their parents were tested by trio whole exome sequencing (trio-WES), sanger sequencing validation sites, and the hazard of their compound heterozygous variants was predicted. Literature review was conducted through domestic and international databases to collect reported RTTN gene mutation cases. 【Results】 Three patients in this family had anomalies of the septum pellucidum, hypoplasia of the corpus callosum and other brain malformations during fetal period. The proband (G2) and fetus (G3) showed intrauterine growth retardation and MCPH in late pregnancy; besides, G2 was born with global developmental delay. Trio-WES detected a c.2101(exon16)C>T(p.Arg701Ter,1526) nonsense and a c.2863(exon22)G>A(p.Glu955Lys)missense in the RTTN gene of G2 and G3, which were inherited from their father and mother, forming a compound heterozygous variant. According to the American College of Medical Genetics and Genomics (ACMG) variant classification guidelines, two variants were likely to be pathogenic (LP) and uncertain significance (VUS). Among them, c.2863(exon22)G>A was a newly discovered missense, which was predicted by the software to be harmful to the gene product. 【Conclusions】 Complex heterozygous variations of RTTN gene (c.2101C>T and c.2863G>A) are the genetic cause of MCPH in this family. This report has enriched the variation spectrum of RTTN gene, provided guidance for prenatal diagnosis and reproduction of this family, as well as material and reference for further understanding of the diseases caused by this gene mutation.