Objective:To identify the genetic variation in a mucopolysaccharidosis type Ⅱ(MPS Ⅱ)family, and conduct a functional study of iduronate-2-sulfatase(IDS): c.323A>C.Methods:A five-generation MPS Ⅱ family of 83 individuals including 4 patients from northern China was collected. Urine mucopolysaccharide and Alder-Reilly body were tested to assist the clinical diagnosis of MPS Ⅱ. IDS enzyme activity was detected on core family members. By the whole exome sequencing of a MPS Ⅱ patient in this family and bioinformatics analysis, the variant was screened and further identified by PCR-Sanger sequencing. Finally, to validate the function of the variant in vitro, the wild-type IDS overexpression plasmid(pCMV-hIDS-WT)and the IDS overexpression plasmid carrying the mutation site(pCMV-hIDS-c.323A>C)were transfected into COS-7 cells and the IDS activity was detected. Results:The proband(Ⅳ3)and Ⅳ4 were diagnosed as MPS Ⅱ by urine mucopolysaccharide, Alder-Reilly body, and IDS enzyme activity tests. Ⅳ3, Ⅳ4, Ⅲ19, and Ⅲ32 were determined to carry IDS: c.323A>C missense variant through the whole-exome sequencing, and diagnosed as MPS Ⅱ. Meanwhile, Ⅱ2, Ⅱ4, Ⅱ8, Ⅱ12, Ⅱ14, Ⅲ5, Ⅲ7, Ⅳ14 in the MPS Ⅱ family carried IDS: c.323A>C missense variant, and were excluded as MPS Ⅱ. The in vitro experiment in COS-7 cells showed that the missense mutation led to a significant decrease in IDS enzyme activity. Conclusion:The variant IDS: c.323A>C: p.Y108S significantly decreases the activity of IDS enzyme in vivo and in vitro, and it is identified as a pathogenic variant for MPS Ⅱ.

We reported a case of mosaic trisomy 2.The patient was a 29-year-old gravida who underwent amniocentesis at 20 weeks of gestation because of high risk of trisomy-21 in the first trimester screening.The test result revealed a karyotype of 47,XN,+2[10]/46,XX[40].At 26 gestational weeks,the fetus was found severe fetal growth restriction and oligohydramnios which was considered to be at risk of mosaic trisomy 2.The pregnancy was terminated at 27+ gestational weeks.The fetus had obviously abnormal appearances,including dolichocephaly,low-set ears,and micromandible.Autopsy was not performed due to the parents' refusal.

OBJECTIVETo assess the detection of maternal serum alpha fetoprotein (MSAFP) and free beta-HCG levels of second trimester for screening of fetal gastroschisis and omphalocele.

METHODSClinical data of 622 639 pregnant women from 5 prenatal screening centers in Hangzhou during October 2007 and September 2016 were analyzed retrospectively. Thirty cases of gastroschisis and 30 cases of omphalocele diagnosed by ultrasonography and postmortem findings were enrolled in the study and 116 cases of pregnant women with normal fetal development during the same period were selected as control group. The cut-off value and area under ROC curve (AUC) of MSAFP and free β-hCG for diagnosis of fetal gastroschisis and omphalocel were analyzed.

RESULTSMSAFP levels of women with fetal gastroschisis and omphalocele were 4.41 (0.88-11.69) MOM and 2.31 (0.72-23.20) MOM, which were significantly higher than that of control group[0.98 (0.41-2.26) MOM, all<0.01]. Free β-hCG level of women with fetal gastroschisis was 1.25 (0.35-19.94) MOM, which was significantly higher than that of control group[0.86 (0.17-6.11) MOM,<0.05). But there were no significant difference in free β-hCG between fetal omphalocele group[1.03(0.21-8.95)]and control group (>0.05). The AUCs of MSAFP for diagnosis of gastroschisis and omphalocele were 0.897 (95%:0.822-0.972) and 0.852(95%:0.762-0.942), respectively (all<0.01). Taking 1.655 MOM as the cut-off value of MSAFP for abdominal wall defects (gastroschisis and omphalocele), the sensitivity was 68.30%, specificity was 99.60% and Youden index was 0.649.

CONCLUSIONSMSAFP of second trimester is a better biomarker than free β-hCG in screening abdominal wall defects.

OBJECTIVETo determine the molecular etiology for a family affected with autosomal dominant polycystic kidney disease and provide prenatal diagnosis for the family.

METHODSClinical data of the family was collected. Target region sequencing with monogenetic disorders capture array combined with Sanger sequencing and bioinformatics analysis were performed in turn. SIFT and NCB1 were used to evaluate the conservation of the gene and pathogenicity of the identified mutation.

RESULTSTarget region sequencing has identified a novel c.11333C to A (p.T3778N) mutation of the PKD1 gene in the proband and the fetus, which was confirmed by Sanger sequencing in three affected individuals from the family. The same mutation was not detected in healthy members of the pedigree. Bioinformatics analysis suggested that the mutation has caused a likely pathogenic amino acid substitution of Threonine by Aspartic acid, and Clustal analysis indicated that the altered amino acid is highly conserved in mammals.

CONCLUSIONA novel mutation of the PKD1 gene has been identified in an affected Chinese family. The mutation is probably responsible for a range of clinical manifestations, for which reliable prenatal diagnosis and genetic counseling may be provided.

Adult ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation ; Pedigree ; Polycystic Kidney, Autosomal Dominant ; genetics ; TRPP Cation Channels ; genetics ; Young Adult

OBJECTIVE To investigate the effect of mono-2-ethylhexyl phthalate(MEHP) on proliferation of primary neural stem cells(NSCs)of rats and NE-4C cells of mice and on the migration of NE-4C cells and the mechanism. METHODS NE-4C or NSCs were treated with MEHP 1,10,100 and 1000 μmol · L-1 for 72 h,respectively. The cytotoxicity was estimated with the cell counting kit-8 (CCK-8). Cell proliferation was analyzed by EdU assay. The mRNA expression levels of the glucocorticoid receptor(GR),signal transducer and activator of transcription 3(Stat3)and sex determining region Y (SRY)-box 2(Sox2) were detected by qRT-PCR. The protein expression levels of total GR,GRβ, Sox2,Stat3 and p-Stat3 were measured by Western blotting. RESULTS Cell viability of NE-4C cells and NSCs at MEHP 1000μmol·L-1 was significantly decreased,which was 70.3%and 40.0%of the control group, respectively. EdU assay showed that MEHP 100 μmol · L-1 decreased NE-4C cells and NSCs by 74.8%and 12.0%(P<0.05)compared with control. The effect of MEHP on the cell migration of NE-4C was evidenced by the fact that the migration was obviously reduced to (63.4±2.0)%(P<0.05)after treatment with MEHP 100μmol · L-1 for 72 h. The mRNA expression levels associated with proliferation and migration in NE-4C of GR,Stat3 and Sox2 in MEHP 100 μmol · L-1 group were down-regulated to 49.8%,26.0% and 14.0%of control(P<0.05). At MEHP 100μmol · L-1,mRNA of GR, Stat3 and Sox2 in NSCs declined to 10.0%,14.0% and 15.3% of normal control. Western blotting results revealed that protein expressions of GR,GRβ,Sox2 and p-Stat3 were remarkably inhibited by MEHP 100 μmol · L-1 in that the relative expression of NE-4C was 0.92 ± 0.17,0.87 ± 0.35,0.81 ± 0.22 and 0.62 ± 0.24(P<0.05). The corresponding protein expression in NSCs was 0.82 ± 0.20,0.56 ± 0.12,0.84 ± 0.36 and 0.53 ± 0.20(P<0.05)when the cells were treated with MEHP 100μmol · L-1 for 72 h. CONCLUSION MEHP can inhibit the proliferation and migration of NE-4C cells and NSCs possibly by decreasing Stat3 and Sox2 that are mediated by GRβ.

Objective To re-evaluate the diagnoses of ankylosing spondylitis (AS) and nonradiographic axial spondyloarthritis (nr-axSpA) and analyze the incidence and reason of misdiagnosis.Methods Patients who were previously diagnosed as AS and nr-axSpA before referrals to Peking Union Medical College Hospital (PUMCH) were re-evaluated by three rheumatologists of PUMCH according to the modified New York criteria for AS and Assessment of SpondyloArthritis international Society (ASAS) axial SpA classification criteria for nr-axSpA.Results Totally 87 prior AS patients and 53 prior nr-axSpA patients were enrolled in this study.After re-evaluation,57 patients were still diagnosed as AS and 16 patients were still diagnosed as nr-axSpA.The misdiagnosis incidences were 34.48％ and 69.81％,respectively.The misdiagnosis incidence of nr-axSpA was higher than that of AS (P ＜ 0.01).Conclusions The misdiagnosis of AS were mainly due to the misjudgment of sacroiliac joints by CT.The misdiagnosis of nr-axSpA were mainly due to the misjudgment of sacroiliac joints by magnetic resonance imaging.Moreover,the misuse of ASAS axial SpA classification criteria contributed to the misdiagnosis also.

Objective To evaluate the clinical value of cardiovascular function in patients with type 2 diabetes mellitus by wave intensity and tissue Doppler imaging.Methods According to the course of disease,30 patients with type 2 diabetes mellitus were divided in to two groups:T2DM group 1,the course of disease between 1 year to 5 years; T2DM group 2,the course of disease was longer than 5 years.30healthy volunteers were considered as control group.The parameters of accelerating wave intensity(W1),decelerating wave intensity(W2),negative area(NA),time interval between the R-wave of ECG and the first peak(R-W1),the first and second peaks (W1-W2),elasticity modulus (Ep),stiffness index (β),arterial compliance(AC),pulse wave velocity(PWVβ) and intima media thickness (IMT) were calculated by wave intensity.The left ventricular ejection fraction (LVEF),left ventricular ejection time (LVET),early diastolic peak speed (E),late diastolic peak velocity (A),E / A and isovolumic relaxation time(IVRT) were detected by two dimensional echocardiography.Mitral annular early diastolic peak speeds (Em),late diastolic peak velocity (Am) and Em / Am were detected by tissue Doppler imaging.Then three groups data were recorded and compared.Results Compared with the control group,β,PWVβ,Ep in T2DM group increased significantly(P ＜0.01 or P ＜0.05),while AC decreased(P ＜0.05).IMT in T2DM group 2 was thicker significantly than the control group(P ＜0.01).PWVβ,Ep,IMT in T2DM group 2 were significantly higher than those of T2DM group 1 (P ＜0.05).W2 in T2DM group 2 was significantly higher than those of T2DM group 1 and the control group(P ＜0.01 or P ＜0.05).E/A,Em and Em/Am in T2DM group were significantly lower than those of the control group (P ＜0.05 or P ＜0.01).Compared with T2DM group 1,Em in T2DM group 2 decreased significantly(P ＜0.05).In T2DM group,there were positive correlation between IMT and Ep,β,PWVβ (r =0.48,r =0.385,r =0.478,P ＜0.05 or P ＜0.01),positive correlation between Em and E (r =0.404,P ＜0.05) ;positive correlation between Em/Am and E/Em,E/A (r =0.602,r =0.559,P ＜0.01) ; negative correlation between Em and IVRT (r =-0.653,P ＜0.01);positive correlation between W1 and LVEF (r =0.627,P ＜0.01); positive correlation between W1-W2 and lVET (r =0.847,P ＜ 0.01) ; no correlation between W2 and E/A,E/Em,Em/Am,IVRT(P ＞0.05).Conclusions The elasticity of carotid artery and the diastolic function of left ventricular decreased in T2DM patients,while the systolic function of left ventricular was normal.

Objective To evaluate the function and synchronization of left atrial in patients with chronic heart failure(CHF) by real-time three dimensional echocardiography (RT-3DE).MethodsThirty healthy controls and thirty-two patients with CHF were studied.The CHF were divided into group CHF1 with the normal of left ventricular filling pressure and group CHF2 with the elevated of that.From the volume-time curve of RT-3DE workstation,the maximum volume (LAVmax),the minimum volume (LAVmin),the volume before contraction (LAVp) of left atrial were acquired.Accordingly,the total,passive and active ejection fraction could be calculated.The synchronization parameters derived from RT3DE were the dispersion and maximum difference of time to minimum volume for left atrial segments (Tmsv- 16-SD,Tmsv 12-SD,Tmsv-6-SD,Tmsv- 16-Dif,Tmsv 12-Dif,Tmsv-6 Dif).The index of volume and time were corrected by the body surface area and interval of R-R,respectively.The correlation were analysed in them.Results ① All the index of volume before and after correction were gradually increasing from control to CHF2 group,but the LATEF,LAPEF and LAAEF were just the reverse,with all parameters had significant difference among total groups( P ＜0.01 ).②The synchronization parameters were significantly higher from control to CHF2 group( P ＜0.01 ).Comparing between two groups,no significant difference of the synchronization parameters were found between CHF1 group and CHF2 group ( P ＞ 0.05),except Tmsv-16-SD％ and Tmsv-16-Dif％ ( P ＜0.01 ).③The Tmsv-16-SD％ and Tmsv-16-Dif％ had a positive correlation with the index of volume and a negative correlation with the ejection fraction.The most significantly correlation parameters was LAVmaxI,with r =0.75 and P ＜0.01.ConclusionsThe function of left atrium are reduced in patients of CHF,there are dyssynchronization.The higher of the filling pressure of left ventricle,the lager of the dyssynchronization of left atrium.RT-3DE may paly important roles in the evaluation of function and synchronization of left atrium.

Seventy eight infertile women were examined by three dimensional hysterosalpingocontrast-sonography(3D-HyCoSy) and X-ray hysterosalingography (HSG) to observe flowing condition of fallopian tube.With the laparoscopy as gold standard,the sensitivity,specificity,positive predictive value and negative predictive value of 3D-HyCoSy in diagnosis of fallopian tube infertility were 94.7％,83.7％,93.9％ and 82.4％ ; those for HSG were 96.4％,82.6％,92.3％,93.0％ and 90.5％,respectively.U test showed that two methods had good consistency. The results suggested that 3D-HyCoSy provides an effective method in evaluation of fallopian tube pathology for infertile women.

ObjectiveTo evaluate whether endometrial blood flow measured by three-dimensional power Doppler ultrasound can predict pregnacy after the treatment of in vitro fertilization and embryo transfer (IVF-ET).MethodsNinty women who received the treatment of IVF-ET examined by threedimensional power Doppler imaging at the day of inject human chorionic gonadotropin(HCG) were enrolled.All patients were divided into pregnancy group (43 cases,include biochemical pregnancy and clinical pregnancy ) and non-pregnancy group(47 cases).The diagnostic value of parameters derived from threedimensional power Doppler imaging was evaluated by using the receiver operating characteristic (ROC)curve.ResultsDifferences of these parameters,including vascularization index (VI),flow index(Fl) and vascularization flow index (VFI)were considered statistically significant between two groups,the parameters of uterine artery and endometrial volume had no significant differences between two groups.The area under ROC curve of VI,FI and VFI were 0.880,0.713 and 0.646,respectively.Among these parameters,VI had the highest diagnostic value for discriminating between benign and malignant endometrium,the cut-off value of which was 0.733 with a sensitivity of 84.2％.Conclusions Threedimensional power Doppler ultrasound was useful for evaluating endometrial blood flow in IVF-ET cycles.Endometrial blood flow parameters may be useful predictors for pregnancy.