Objective:To compare the genotypes and phenotypes between the monozygotic twins via whole genome sequencing to further clarify the autosomal dominant inherited neurofibromatosis type 1 (NF1) variants related to congenital pseudarthrosis (CP).Methods:According to the diagnostic criteria of congenital tibial pseudarthrosis and the clinical diagnostic criteria of NF1, two pairs of monozygotic twins with NF1 were included. Both were female and only one of each pair had congenital pseudarthrosis. The other did not have congenital pseudarthrosis. Whole genome sequencing was performed using the peripheral blood of the two pairs of monozygotic twins. Customized bioinformatics analysis was then performed to identify single nucleotide variants (SNVs), short insertion deletion variants (InDel), copy number variants (CNVs), and structural variants (SVs). Classified the variants according to the American College of Medical Genetics and Genomics (ACMG) and ClinGen criteria. The germline variants within the monozygotic twins were compared to identify the CP patients' unique variants. The shared pathogenic or likely pathogenic germline variants between the unique variants in the CP patients from the twins were also analyzed. Further, the identified disease-causing variants were validated by Sanger sequencing in the family of the twins and their parents. Finally, the genotypes and phenotypes regarding the pathogenic variants of the NF1 gene among the twins were characterized. Results:Both the two monozygotic twins were identified pathogenic variants in the NF1 gene. One with c.3047_3048del (p.Cys1016SerfsTer4), and the other with c.4267A>G (p.Lys1423Glu). By Sanger sequencing validation in family quads, the two CP patients and their siblings harbored de novo heterozygous variants of the NF1 gene. In addition to the NF1 gene, no other genes were identified pathogenic or likely pathogenic variants uniquely in the CP patients compared with their twin sisters, as well as SVs and CNVs. In addition, by analyzing the rare and damaging variants in the two CP patients from the two twins, they had no overlapping genes against the SNVs, InDels, SVs, or CNVs. Conclusion:Whole genome sequencing revealed that both the two monozygotic twins with NF1 were detected pathogenic variants of gene NF1. No other pathogenic variants specific to the CP patients among the twins were identified. The two CP patients shared no other common genes from the detected likely pathogenic variants.

Objective To explore the changes in gravida's age and its influences on maternal and neonatal complications under China's two-child policy.Methods This study retrospectively analyzed the clinical data such as adverse gestational complications and fetal condition of 42 771 gravidas delivering at Changzhou Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University from July 2013 to December 2017.According to their age at delivery,they were divided into three groups:the younger maternal age group (1 140 cases,＜20 years),the advanced maternal age group (4 307 cases,≥ 35 years) and the median maternal age group (37 324 cases,≥ 20 and ＜35 years).Chi-square test was used to compare the differences among groups.Cochran-Armitage test was used for trend analysis.The risks of various complications in younger and advanced maternal age groups were analyzed by binary logistic regression analysis.Results (1) The proportion of advanced maternal age pregnancies tended to rise gradually year by year (Z=-9.909,P＜0.001).However,the figure of younger gravidas remained low and presented a downward trend (Z=10.685,P＜0.001).(2) The incidence of pregnant complications in the younger,advanced and the median maternal age groups were 52.8％ (602/1 140),72.3％ (3 116/4 307) and 56.5％ (21 091/37 324),respectively.Compared with the median maternal age group,the advanced maternal age group was at greater risks of premature delivery [9.0％ (3 343/37 324) vs 11.6％ (499/4 307),x2=124.233,P＜0.001],fetal growth restriction (FGR) [0.6％ (218/37 324) vs 1.2％ (50/4 307),x2=20.087,P＜0.001],postpartum hemorrhage [5.7％ (2 120/37 324) vs 7.8％ (336/4 307),x2=31.299,P＜0.05],hypertensive disorders in pregnancy(HDP) [4.2％ (1 561/37 324) vs 8.7％ (376/4 307),x2=180.013,P＜0.001],gestational diabetes mellitus (GDM) [7.6％ (2 845/37 324) vs 15.1％ (650/4 307),x2=280.126,P＜0.001]and placenta previa [1.7％ (621/37 324) vs 3.8％ (165/4 307),x2=97.904,P＜0.001],and the younger maternal age group was at greater risks of HDP [4.2％ (1 561/37 324) vs 5.9％ (67/1 140),x2=4.234,P=0.040],fetal distress [3.5％ (1 325/37 324) vs 5.1％ (58/1 140),x2=7.546,P=0.006],premature delivery [9.0％ (3 343/37 324) vs 15.0％ (171/1 140),22=48.668,P＜0.001] and FGR [0.6％ (218/37 324) vs 1.1％ (12/1 140),x2=4.086,P=0.043].(3) Gestational complications in the younger maternal age group were mainly related to the fetuses such as premature rupture of membranes (PROM) and premature delivery,while the advanced maternal age group had a higher incidence of maternal complications,especially GDM and HDP.(4) Most of the gravidas of advanced maternal age with HDP developed severe preeclampsia (47.9％,180/376),while mild preeclampsia was dominant in the median maternal aged HDP women (45.4％,708/1 561).(5) The advanced maternal age group had higher risk of stillbirth,premature delivery,FGR,placenta previa,GDM,HDP and postpartum hemorrhage [OR(95％CI):1.91 (1.29-2.84),1.33 (1.21-1.46),1.66 (1.21-2.28),2.56 (2.15-3.04),2.39 (2.19-2.61),2.36 (2.11-2.65),1.46 (1.31-1.62);all P＜0.05],but lower risks of fetal distress and PROM [OR(95％CI):0.79 (0.65-0.95) and 0.88 (0.81-0.96);both P＜0.05].The younger maternal age group had a higher risk of premature delivery [OR(95％CI):1.97 (1.61-2.40);P＜0.001],but significant lower risks of PROM and GDM [OR(95％CI):0.77 (0.62-0.95) and 0.05 (0.02-0.16);both P＜0.05].Conclusions Maternal age is closely related to the adverse outcomes of pregnancy.Two-child policy in China will bring about changes in maternal age and composition of pregnant complications.

Objective To observe the effect of high frequency repetitive transcranial magnetic stimulation ( rTMS) on executive function in patients after stroke .Methods Thirty-six stroke patients with executive dysfunction in the Second Affiliated Hospital of Guangzhou Medical University from March 2016 to April 2017 were randomly divided into experimental group ( n=18 ) and control group ( n=18 ) .The experimental group received 10 Hz rTMS of the left dorsolateral prefrontal cortex ( DLPFC ) and cognitive functional training , while the control group received sham rTMS and cognitive functional training , for four weeks.Mini-Mental State Examination(MMSE), Wisconsin Card Classification Test(WCST), Digit Symbol Test( DST) and Digital Span ( DS) were used for cognitive assessment at baseline and therapy completed . Results After a four weeks therapy , the total number of responses showed no significant difference in WCST between two groups.But the control group after treatment showed that MMSE score (17.44 ±4.73) and DST score (4.50 (2.75, 8.25)) were improved compared with that before treatment (MMSE score 15.28 ±4.61, t=-8.371, P=0.000;DST score 4.00 (2.75, 7.25), Z=-2.122, P=0.049), and the DS score and the numbers of correction , classification and continuous errors of WCST showed no significant difference compared with that before treatment .The experimental group after treatment showed that MMSE score (20.67 ±4.59), DST score (7.50(4.75, 12.50)), DS score (recite in order 7.00 (7.00, 8.00), recite in reverse order 3.00 (2.00, 4.00)), the numbers of correction of WCST (24.61 ± 8.30), the numbers of continuous errors (12.17 ±5.08), the numbers of classification (2.00 (2.00, 3.00) were improved compared with that before treatment ( MMSE score 15.50 ±5.24, t=-21.013, P=0.000;DST score 3.00(1.00, 7.00), Z=-3.757, P=0.000; DS score recited in order 6.00(5.00, 6.00), Z=-3.703, P=0.000;DS score recited in reverse order 2.00(1.00, 3.00), Z=-3.494, P=0.000;The numbers of correction of WCST (16.50 ±9.34), t=-6.544, P=0.000); The numbers of continuous errors (18.06 ±5.63, t=9.744, P=0.000); The numbers of classification (1.00 (1.00, 2.00), Z=-3.900, P=0.000 ).And the curative effect was better than that of control group . Conclusion High frequency rTMS combined with cognitive function training can improve the executive dysfunction of stroke patients , and the improvement is better than cognitive functional training alone .

Objective To study the effects of the rehabilitation robot-assisted task-oriented training on the hand function in patients after stroke. Methods From June, 2015 to September, 2016, 35 inpatients suffering from stroke were randomly allocated to control group (n=17) and trial group (n=18). Based on the routine rehabilitation, the trial group accepted robot-assisted task-oriented training, while the control group accepted therapist-assisted task-oriented training, for two weeks. They were measured the active range of motion (AROM) of fingers, assessed with fingers motor of Fugl-Meyer Assessment (FMA) and modified Barthel Index (MBI) invovled with hands before and after train-ing. Results The inpatients dropped three in the control group, two in the trial group. AROM of extension and flexion of all the fingers, the AROM of extension and total of three fingers of thumb, index and middle, and the total AROM of each finger improved in the trial group af-ter training (t>2.937, P<0.05), while the AROM of extension and flexion of all the fingers, AROM of extension, flexion and total of the fin-gers of thumb, index and middle, total AROM of the fingers of thumb, index and little improved in the control group after training (t>2.528, P<0.05);the AROM of extension and total of the fingers of thumb, index and middle, and the total AROM of fingers of thumb and index im-proved more in the trial group than in the control group (t>2.535, P<0.05). The scores of mass flexion, mass extension, opposition, cylinder grip, spherical grip and total score of FMA improved in the trial group after training (Z>2.000, P<0.05), while the scores of mass extension, opposition and the total score of FMA improved in the control group after training (Z>2.000, P<0.05). There was no significant difference between the two groups on the items and total scores after training (P>0.05). The scores of feeding, dressing, toilet transfers, bathing, groom-ing of MBI and the total score of them improved in the trial group after training (Z>2.041, P<0.05), while the total score of MBI improved in the control group after training (Z=-2.527, P<0.05). There was no significant difference between the two groups in the items and total scores after training (P>0.05). Conclusion The rehabilitation robot-assisted task-oriented training can improve AROM of hemiplegic fingers and grip function.

Objective To observe the effect of virtual reality robotic hand on hand motor function and activities of daily living of pa-tients after stroke. Methods From June, 2015 to June, 2016, 32 patients with hand motor dysfunction were assigned into experimental group (n=16) and control group (n=16). The experimental group received training with virtual reality robotic hand and hand based rehabilitation, while the control group received hand based rehabilitation only, for four weeks. They were evaluated with Fugl-Meyer Assessment (FMA) of fingers and wrists and modified Barthel index (MBI) before and after treatment. Results The total score and the scores of items of FMA and MBI improved after treatment in the experimental group (Z>3.45 or t>3.45, P<0.01). The total score and the scores of the finger, wrist of FMA, and the scores of the eating, dressing and grooming of MBI improved in the control group (Z>2.07 or t>4.18, P<0.05). The total scores and scores of the items of FMA and MBI improved more in the experimental group than in the control group (Z>2.14 or t>3.20, P<0.05). Conclusion Virtual reality robotic hand training can promote the recovery of hand function and activities of daily living in patients af-ter stroke.

OBJECTIVETo assess the association of single nucleotide polymorphisms (SNPs) of NLRP3 gene with metabolic syndrome (MetS).

METHODSA total of 885 subjects including 410 MetS patients and 475 healthy controls were recruited. MetS was defined based on the National Cholesterol Education Program in Adult Treatment Panel III criteria. Two common SNPs of the NLRP3 gene, rs10754558 and rs4612666, were detected using polymerase chain reaction-restriction fragment length polymorphism method.

RESULTSThe frequencies of G allele and GG genotype of the NLRP3 rs10754558 in the MetS group were significantly higher than those of the control group. Logistic regression analysis showed that the GG genotype (OR=2.223, 95%CI: 1.296-6.924, P=0.00034) and G allele (OR=1.440, 95%CI: 1.189-4.063, P=0.00028) were associated with increased risk of MetS. NLRP3 rs10754558 GG genotype was associated with higher level of insulin resistance and visceral adiposity index. No association of NLRP3 rs4612666 SNPs with susceptibility to MetS was identified in this population.

CONCLUSIONNLRP3 gene rs10754558 polymorphisms are associated with increased risk of MetS. The G allele and genotype GG are risk factors for MetS.

Adult ; Aged ; Carrier Proteins ; genetics ; Female ; Genotype ; Humans ; Insulin Resistance ; Logistic Models ; Male ; Metabolic Syndrome ; etiology ; genetics ; Middle Aged ; NLR Family, Pyrin Domain-Containing 3 Protein ; Polymorphism, Single Nucleotide

Objective To investigate the relationship between the single nucleotide polymorphisms(SNPs) of disrupted in schizophrenia 1 (DISC1) gene and autism in Chinese Han children.Methods Genome-wide SNP genotyping was performed by using Illumina HumanHap CNV370-Duo Chip in 278 autistic trios,157 autistic individuals and 435 healthy controls.Genotype data of SNPs within DISC1 gene were selected.The association between these SNPs loci and autism was analyzed through case-control association analysis and family-based transmission disequilibrium test.Results Fifty-two SNPs were involved for further analysis.1.Case-control association analysis showed that 6 SNPs (rs4658939,rs2793093,rs10495309,rs2492367,rs1 1122362,rs1073179) and 7 haplotypes (AGAAAG constructed with rs823163-rs823161-rs4658933-rs1417585-rs10864693-rs6541281,GGG and AAA constructed with rs4658939-rs2793093-rs10495309,GG and AG constructed with rs2492367-rs12046794,GAA constructed with rs9432040-rs2356606-rs1 1122362 and GG constructed with rs9431714-rs1073179) had significant differences between autistic patients and controls(x2 =4.704,4.915,5.568,8.087,4.043,5.183,5.369,5.295,4.440,5.304,7.615,4.018,4.811,P =0.030,0.027,0.018,0.005,0.044,0.023,0.021,0.021,0.035,0.021,0.006,0.045,0.028).2.In the transmission disequilibrium test analysis,2 SNPs (rs4658945,rs11122362) and 2 haplotypes (AG constructed with rs10495310-rs4658945,GAA constructed with rs9432040-rs2356606-rs11122362) showed significant transmission disequilibrium(x2=4.445,5.400,3.973,5.126,P =0.035,0.020,0.046,0.024).Conclusions The polymorphism of rs11122362 and GAA haplotype constructed with rs9432040-rs2356606-rs11122362 are associated with autism,and DISC1 gene is a susceptibility gene for autism in Chinese Han children.

Objective:Mutation in the gap junction beta 6 (GJB6) gene has been reported to be associated with an autosomal dominant disorder hidrotic ectodermal dysplasia (HED), characterized by congenital nail clubbing, alopecia and palmoplantar keratoderma. The aim of this study is to investigate relationship between genetic mutation in GJB6 and HED in an affected Chinese family.
Methods:We selected a Chinese HED family consisting of a total of 17 individuals including 8 HED patients (5 males and 3 females). The whole coding region of GJB6 was amplified by polymerase chain reaction and sequenced.
Results:Sequence analysis identified a heterozygous missense mutation c.31G>A (p.G11R) in GJB6 gene of affected individuals, but not in healthy individuals.
Conclusion:A c.31G>A (p.G11R) missense mutation in GJB6 gene is the genotypic characteristic for HED in Chinese population.

Autism is a group of etiology and clinical heterogeneous neurodevelopmental disorders with an onset before 3 years old. It has 3 core characteristics: deficits in verbal communication; impairment of social interaction; restricted interests and repetitive behaviors. The incidence is increasing over time worldwide. Twin and family studies have demonstrated that autism has a high heritability (>90%). Although certain progress of autism genetic study has been made in the last decades and several autism susceptibility genes and loci have been identified, there are still about 70%-80% of patients for whom an autism-related genetic change cannot be identified.
Autistic Disorder ; genetics ; Child, Preschool ; Epigenomics ; Genetic Heterogeneity ; Humans ; Infant

Objective To study the clinical characteristics and genetic cause of a Chinese family affected with paroxysmal kinesigenic dystonia(PKD).Methods The detailed clinical data and the blood samples of the affected patients with PKD and their relatives were collected.After genomic DNA was extracted from blood leukocytes,target linkage analysis Was performed using multiplex PCR by microsatellite marker's located in the reported critical region on chromosome 16.All exons and flanking regions of SCNN1G and ITGAL genes were amplified by PCR-sequence.Results In this three-generation 12 member family,5 individuals have been diagnosed as PKD.Target linkage analysis suggested the disease gene linked to chromosome 16.between D16S3396 and D16S3057 with two-point LOD score of 1.47 at recombination fraction(θ)=0.0.All affected individuals shared a common haplotype which co-segregated with the phenotype.Except for 8 reported SNPs,no pathologic sequence variants were found in candidate genes SCNN1G and ITGAL.Conclusions The studied family is genetically linked to the reported critical locus of PKD on chromosome 16.SCNN1G and ITGAL were ruled out as the causative genes for the studied pedigree.Further genetic analysis in this family may reveal new genetic cause responsible for PKD.