OBJECTIVE: To determine the carrier rate for 21 inherited metabolic diseases among a Chinese population of childbearing age. METHODS: A total of 897 unrelated healthy individuals (including 143 couples) were recruited, and DNA was extracted from their peripheral blood samples. Whole exome sequencing (WES) was carried out to screen potential variants among 54 genes associated with 21 inherited metabolic diseases. Pathogenic and likely pathogenic variants and unreported loss-of-function variants were analyzed. RESULTS: One hundred fourty types of pathogenic/likely pathogenic variants (with an overall number of 183) and unreported loss-of-function variants were detected, which yield a frequency of 0.20 per capita. A husband and wife were both found to carry pathogenic variants of the SLC25A13 gene and have given birth to a healthy baby with the aid of preimplantation genetic diagnosis. The detected variants have involved 40 genes, with the most common ones including ATP7B, SLC25A13, PAH, CBS and MMACHC. Based on the Hardy-Weinberg equilibrium, the incidence of the 21 inherited metabolic diseases in the population was approximately 1/1100, with the five diseases with higher incidence including citrullinemia, methylmalonic acidemia, Wilson disease, glycogen storage disease, and phenylketonuria. CONCLUSION This study has preliminarily determined the carrier rate and incidence of 21 inherited metabolic diseases among a Chinese population of childbearing age, which has provided valuable information for the design of neonatal screening program for inherited metabolic diseases. Pre-conception carrier screening can provide an important measure for the prevention of transmission of Mendelian disorders in the population.
Asians/genetics* ; China ; Exome ; Female ; Humans ; Infant, Newborn ; Metabolic Diseases/genetics* ; Mitochondrial Membrane Transport Proteins/genetics* ; Oxidoreductases/genetics* ; Whole Exome Sequencing

Objective To study the value of score for neonatal acute physiology Ⅱ(SNAP]Ⅱ) and its extension version Ⅱ (SNAPPE-Ⅱ) in predicting neonatal necrotizing enterocolitis (NEC) outcome.Methods We explored 73 NEC patients by statistics who were treated in our hospital from January 2002 to January 2012.The patients were divided into two groups:surgery group and non-surgery group,then they were divided into subgroups:alive group and death group.The general information including birth weight,age,clinical manifestations,treatment of patients were collected.Every patient was checked and scored by the methods SNAP-Ⅱ] and SNAPPE-Ⅱ in time.Results The scores (27.0 ± 2.3,26.5 ± 1.8) of surgery group including SNAP-Ⅱ and SNAPPE-Ⅱ were higher than those (14.0 ± 2.1,15.0 ± 2.5) in the non-surgery group(P ＜ 0.01).The scores(31.0 ± 3.2,31.0 ± 3.4) of the death group including SNAP-Ⅱ and SNAPPE-Ⅱ were higher than those(11.0 ± 2.5,10.0 ± 3.6) in the alive group(P ＜ 0.01).According to the area under the curve(AUC) analyzed by the receiver operating characteristic(ROC) curve for measuring the scores of surgery predicting,AUC was 0.726 for SNAP-Ⅱ and 0.732 for SNAPPE-Ⅱ.The value of predicting surgery risk was 20 and 24 respectively.According to the AUC analyzed by the ROC curve for measuring the scores for surgery predicting,AUC was 0.752 for SNAP-Ⅱ and 0.825 for SNAPPE-Ⅱ.The value of predicting mortality risk was 31 and 33 respectively.All P values were less than 0.01 and there were significant differences.Conclusion The two kinds of score for neonatal acute physiology have an important significance in predicting surgery and mortality risk of NEC.

Objective To study the relationship between umbilical cord leptin levels and fetal growth as well as neonatal birth weight. Methods One hundred and forty - two neonates selected from February 2009 to June 2013 in Shangqiu First People's Hospital according to the different gestational age and birth weight were divided into 3 groups. Group A included 44 cases(small for gestational age,birth weight below the average weight of the 10th percentile at the same gestational age),23 boy cases,21 girl cases;group B included 56 cases(appropriate for gestational age,birth weight at the average weight of the 10th to 90th percentile at the same gestational age),30 boy cases,26 girl cases;group C included 42 cases(large for gestational age,birth weight above the average weight of the 90th percentile at the same gestational age),22 boy cases,20 girl cases. Neonatal body mass index,birth weight,placenta weight and umbilical lep-tin levels of three groups were compared. Results Neonatal birth weight,neonatal body length,body mass index and the placenta weight leptin levels in group A were significantly lower than those of group B,having statistically significant difference(all P ﹤ 0. 001);Neonatal birth weight,neonatal body length,body mass index and the placenta weight leptin levels in group C were significantly higher than those in group B,with statistically significant difference( all P ﹤0. 001). Neonatal birth weight in the boy group was obviously higher than that of the girl group,and the difference was statistically significant(P ﹤ 0. 001). Neonatal leptin levels in the boy group were significantly lower than that of the girl group,and the difference was statistically significant(P ﹤ 0. 001). There were positive correlations between the umbili-cal cord leptin levels and the neonatal birth weight,neonatal length,neonatal weight index and the placenta weight(r =0. 382,0. 276,0. 358,0. 412,all P ﹤ 0. 01). Conclusions The umbilical cord leptin levels are closely associated with neonatal birth weight and intrauterine growth retardation,and it can be used as one of the important indicators for reflec-ting neonatal birth weight and fetal growth.

Objective:This study aimed to explore the effect of Yes-associated protein (YAP) on the growth of the human glioma cell line LN229. Methods:YAPsiRNA was transfected into LN229 cells to knock down the YAP expression. The downregulation of the YAP expression was identified through Western Blot analysis. Colorimetric assay using methyl-thiazolyl-tetrazolium was applied to evaluate cell proliferation ability. Cell invasive activity was examined using Transwell assay. Flow cytometry and AnnexinV were used to detect cell cycle and apoptosis, respectively. The relevant molecules regulating proliferation, invasion, cell cycle progression, and apoptosis were examined through Western Blot analysis. Results:The YAP expression was downregulated after YAPsiRNA was trans-fected into LN229 glioma cells. Reduced YAP expression could arrest the cell cycle at G0/G1 phase, inhibit cell proliferation and inva-sion, and promote apoptosis. The expression of the proliferating cell nuclear antigen (Ki-67), matrix metallopeptidase-9 (MMP-9), cy-clin D1, and Bcl-2 were downregulated. Conclusion:The downregulation of YAP in LN229 cells suppresses cell proliferation and inva-sion, as well as promotes cell apoptosis. This study provides a novel evidence for further study on Hippo-YAP signal pathway in molec-ular pathology of glioma.

ObjectiveTo explore the best time for replacing the long-term indwelling catheter in patients with silicone catheter.MethodsDomestic literature were retrieved and analyzed using Cochrane systematic review,and the meta-analysis of randomized controlled trials was conducted in accordance with inclusion criteria and exclusion criteria so as to look into the most suitable time for replacing the indwelling catheter for patients with silicone catheter at clinical trials.The replacement frequency for the catheters of different size together with the relevant infections in urinary tract infection and relative risk(RR)were used as values for effectiveness of interventions.ResultsA total of 11 literature were retrieved. Meta-analysis results suggested that the RR values of urinary tract infections when the catheters were replaced once every two weeks vs. every four weeks,and once every three weeks vs. every four weeks were 0.51[95% CI(0.40,0.66),P <0.001],0.79[95% CI(0.58,1.08),P = 0.14],respectively.The urinary infection rate of replacing a silicone cathether every 2 weeks was higher than that of every 4 weeks,but there was no difference between that of every 3 weeks and 4 weeks.Conclusion According to the nature of silicone catheter material as well as the clinical indexes, it is most reliable to replace a silicone catheter every four weeks to reach a best clinical outcome.

Objective To investigate the effect of knocking-down Yes-associated protein (YAP)on the biologi-cal characteristics of SNB19 glioblastoma cell.Methods The expression of YAB in SNB19 was knockdown by YAB small interfering RNA (YABsiRNA).The downregulation of YAP expression was identified by Western blot analysis. The proliferative ability of cell was determined by methyl thiazoyl terazolium (MTT).The invasive ability of cell was examined by Transwell assay.Flow cytometry and Annexin V staining were used to detect the cell cycle and apoptosis respectively.The results were analyzed by the statistical software SPSS18.0.Results The expression of YAP in the cells transfected with YAPsiRNA was significantly reduced.The cell proliferation activity of SNB19 cells was inhibited, which decreased from (100.00 ±0.00)%to (52.32 ±3.10)%(F=33.00,P<0.01).The cell cycle was arrested in G0-G1 phase (F=8.76,P<0.01).The cell invasive ability was attenuated apparently,which decreased from (163.20 ±10.10)to (37.71 ±2.52)(F=282.05,P<0.01).The apoptosis ratio of the tumor cell which transfected with YAPsi-RNA was increased from (3.56 ±0.35)%to (18.99 ±0.66)%,(F=931.99,P<0.01).Conclusion Knocking-down YAP expression in glioma cells could inhibit the proliferative activity and invasive ability of SNB19 cell and could induce cell apoptosis.YAP could be served as a potential target for the gene therapy of glioma.

Objective To investigate the inhibitory effect of Notch2 gene on the growth of A172 glioblastoma cells and its underlying mechanism.Methods Blank control group,empty vector transfection group (pcDNA3) and Notch2 plasmid transfection group (Notch2-pcDNA3) were employed in our study; A172 glioma cells in the later two groups were transfected with blank load plasmid and Notch2 plasmid,respectively.Cell growth was detected by MTT assay,and cell cycle and apoptosis were analyzed by flow cytometry.Transwell assay was used to study the cell invasion,and the expressions of important signaling pathway members,including Notch2,epidermal growth factor receptor (EGFR),phospho-Akt (P-Akt),phosphatidylinositol 3-kinase (PI3K),phosphatase and tensin homologue deleted on chromosome 10 (PTEN),caspase3,nuclear factor κB (NF-κB),proliferating cell nuclear antigen (PCNA),intedeukin-2 (Bcl-2),matrix metalloproteinase-2 (MMP-2),MMP-9andcyclin-dependent kinase inhibitor 1 (Cyclin D1),were detected by Western blotting.Results MTT assay showed that the cell viability in the A172 cells of Notch2-pcDNA3 group was significantly decreased as compared with that in the blank control cell and pcDNA3 groups (P＜0.05); decreased cell number at S phase and G2/M phase was noted in A172 cells of Notch2-pcDNA3 group,while cell number at G0/G1 phase increased by 22.1％ as compared with that in the blank control group; the percentage of apoptotic cells in the A172 cells of Notch2-pcDNA3 group (14.8％) was significantly increased as compared with that in the blank control cells (1.2％); the number of invaded cells in the A172 cells of Notch2-pcDNA3 group (17.43) was significantly decreased as compared with that in the blank control cells (35.23).The important members related to PI3K-AKT signaling pathway,including EGFR,p-AKT,PI3K,NF-κB,PCNA,Bcl-2,MMP-2,MMP-9 and Cyclin D1 had obvious decreased expression,while PTEN and Caspase-3,the antagonistic PI3K/AKT activity tumor suppressor proteins,had obvious increased expression.Conclusion The suppressive effect of Notch2 gene on glioma cell proliferation may result from the inhibition of EGFR/PI3K/AKT pathway.

Objective To study the inhibitory effect of knocking down miR-30a-5p on the U87 human glioma xenograft growth and its possible mechanism.Methods Nude mice bearing subcutaneous U87 human glioblastoma were established and separated into three groups (eight for each group) by randomized digital table method,including control group,scr-ODN treated group and AS-miR-30a-5p treated group.After relevant subcutaneous injection treatment,tumor size was measured every other day until the observation period ended.Researchers executed the animals after the treatment,stripped tumor tissues and extracted RNA and protein.Real-time PCR was conducted to detect the expression of miR-30a-5p.The histopathological characteristics and proliferation and apoptosis biological characters (including SEPT7,PCNA,cyclin D1,MMP-2,apoptosis related factor P53,bcl-2 and caspase3) were evaluated by HE and immunohistochemical staining,Westem blot analysis respectively,and the cell apoptosis was detected by TUNEL method.Results In AS-miR-30a-5p treated group,the tumor growth was delayed and the final tumor volume was smaller than that in the control and scr-ODN treated group (F =7.167,P ＜0.05),and the expression of miR-30a-5p was knocked down.The expression of PCNA,cyclin D1 were significantly downregulated while P53,SEPT7 and caspase3 up-regulated.Apoptotic index was increased significantly.Conclusion As-miR-30a-5p suppresses the growth of U87 human gliomas xenografts significantly.Malignant phenotype of tumors are reversed to a considerable degree.Therefore,miR-30a-5p can be a candidate for targeted therapy of human glioma.

ObjectiveTo confirm the effect of miR-93 inhibitor in glioma cell growth and invasion.MethodsMalignant glioma cells were transfected with miR-93 inhibitor by lipofectamin to downregulate their overexpression of miR-93.Real time-PCR was taken to measure miR-93 expression after transfection.The cell cycle kinetics and cell growth rate were detected by flowcytometry and MTT assay,the cell proliferative ability was evaluated by soft agar assay,and the invasive ability was detected by transwell assay.ResultsThe highlevel expression of miR-93 was downregulated effectively in glioma cells after transfecting the miR-93 inhibitor.Meanwhile,the cell cycle progress was delayed,S phase cells were reduced,the speed of growth was slowed,cloning formation ability was receded,the number of cells through the matrigel was reduced,and invasive ability was significantly repressed.ConclusionDownregulation of miR-93 expression could inhibit the proliferative ability and invasive ability of glioma cells.

objective To identify SEPT7as one of the target genes of miR-19a and miR-19b.MethodsmiR-19a inhibitor and miR-19b inhibitor mediated by lipofectamine2000,were transfected to SNB19 glioma cells for knocking down miR-19a/19b overexpression.Real time PCR was conducted to detect the expression of miR-19a/miR-19b in transfected cells.The expression of SEPT7was determined by Western blot analysis.RT-PCR was used to detect the mRNA expression of SEPT7; and Luciferase reporter assay was used to identify the direct regulation of miR-19a/19b on SEPT7.ResultsIn SNB19 glioma cells transfected with miR-19a/19b inhibitor,the expression of miR-19a/miR-19b was significantly reduced,whereas the protein expression of SEPT7 was upreguhtted; no significant change of SEPT7 mRNA level was found and luciferase activity became stronger as compared to control cells.ConclusionSEPT7 is the target negatively regulated by miR-19a and miR-19b.