Objective To evaluate the influence of pregnancy outcomes with different dosage of estradiol valerate in addition to progesterone for luteal supplementation in IVF-ET cycles. Methods From February 2011 to October 2012, 362 patients undergoing IVF-ET in the second affiliated hospital of Kunming Medical University were selected. The patients were randomly assigned into three groups. Since oocyte retrieval, all of them were injected luteal supplementation with progesterone in oil 60 mg/d, im. Group A as control group (112 cases) did not add with pentanoic acid estradiol;Group B for the experimental group (122 cases), added pentanoic acid estradiol 4 mg/d after embryo transfer;Group C for the experimental group (128 cases) added pentanoic acid estradiol 6 mg/d after embryo transfer. Finally the pregnancy outcome and implantation rate were compared for each group. Results There were no significant differences between group A and group B, group B and group C in the rate of embryos grow and pregnancy rate ( >0.05) . The embryo planting rate and pregnancy rate have significant difference between Group A and group C ( < 0.05) . Conclusion 6 mg/d fill estradiol valerate for luteal supplementation may improve implantation rate and pregnancy rate after IVF.

Objective To explore the related factors of fertilization failure after in-vitro fertilization and embryo transfer. Methods 150 patients were divided into total fertilization failure (TFF) group, low fertilization (LFR) group, and control (NFR) group according to fertilization rate. Semen was collected from the male pa-tients; the number, concentration, shape, and progressive motility of sperms were measured. Level of gACE was detected by Western blot. Logistic regression was used to explore the factors affecting fertilization rate. Results The fertilization rate and the concentration , progressive motility , and shape of sperms in were lower TFF group than in LFR group and NFR group (P < 0.05). Western blot proved that level of gACE group was higher in TFF than in LFR group and NFR group (P < 0.05). Logistic regression showed that the fertilization rate, the concen-tration , progressive motility , shape of sperms , and the level of gACE were all the independent risk factors for fertilization failure. Conclusions The concentration, progressive motility, and shape of sperms have impact on IVF. A lower expression of gACE in patients with lower fertilization rate can be used as a potential biomarker for predicting fertilization failure.

Objective To investigate the genetic basis of the children with growth retardation. Methods From January to October 2013, the 56 patients with growth retardation were enrolled in this study. Genomic DNA was extracted from peripheral blood and was analyzed with gene array chips. Results Abnormalities were found in 12 patients (6 cases of sex chromosome abnormalities and 6 cases of autosomal aberration) and the detection rate was 21.4%. Four patients had the copy-number variations of smaller than 2.5Mb in size which could not be found by karyotyping analysis. Conclusions SNP-array gene chip could be used in the genetic diagnosis of growth retardation.

Objective To analyze blood Met、 Phe 、Tyr、 Arg、 Cit、 Orn、 Ser、 Thr、 C0、 C2、 C3、 C14、C14 ∶ 1 , C16 , C16 ∶ 1 , C18 , C18 ∶ 1 and urine 4-OH-PHPLA , 4-OH-PHPPA level of NICCD patient and discuss the application value of diagnosis NICCD. Methods From May 2011 to May 2015, 21 NICCD patient were diagnose in Guangxi Newborn Screening Center. Meanwhile, 100 normal children were selected as the control group. Blood Met, Phe, Tyr and other factors and urine 4-OH-PHPLA, 4-OH-PHPPA level were analyzed by SPSS. Results In the experimental group, blood Met, Phe, Tyr and many other indexes and urine 4-OH-PHPLA, 4-OH-PHPPA level were higher and blood Orn/Cit were lower than the control group(P < 0.05), while blood C2and Cit/Arg were increased (P > 0.05). Conclusion NICCD patient has abnormal biochemical index. Blood test by TMS and urine test by GC-MS are very important in NICCD diagnosis.

Preeclampsia, a serious complication in pregnancy, is a "placental disease" in obstetrics. The growth and development of human and other mammalian embryos are closely related to the placenta. This review summarizes several models for studying preeclampsia including in vitro cell models, placental explants, organoid models, rodent models and primates models, contributing to a deeper understanding of placental development and related pregnancy disorders.

Objective To investigate the genetic basis of patients with intellectual disability,and to assess the application of single nucleotide polymorphisms (SNP)-array in the molecular diagnosis of intellectual disability.Methods Sixty-four patients with intellectual disability who were identified in Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from January 2013 to June of 2015 were enrolled.Genomic DNA was extracted from peripheral blood and was analyzed with Illumina Humancyto SNP-12 300K gene array chip.All identified copy number variants (CNVs) were analyzed with references from databases such as ClinVar,DECIPHER,OMIM and DGV(Database of Genomic Variants),as well as comprehensive literature review from PubMed database to determine the pathogenicity of CNVs.Results Sixteen cases of the above 64 patients were found to have CNVs with genomic alterations,including 6 cases microdeletions/microduplications associated with known syndromes,3 cases microdeletions and microduplications with clear clinical relevance (non-syndrome),1 case numerical chromosome aberration,1 case unbalanced translocation and 5 cases CNVs of unknown clinical significance.The detection rate was 25% (16/64 cases).Among these 16 abnormalities,6 cases of them could not be detected by using karyotyping analysis because their sizes were less than 5 Mb,and the smallest detected missing fragment was 0.53 Mb.Conclusion SNP-array gene chip technique with the advantages of higher efficiency,high-resolution and good accuracy,which can be applied to the genetic diagnosis of intellectual disability.

OBJECTIVE: To identify and analysis three ABO variant Bw subtypes. METHODS: Serological assays were carried out to identify the ABO blood group of the proband. ABO gene was identified by Sanger sequencing. RESULTS: The genotype of three individuals are ABO*Bw.11/0.01.02, ABO*Bw.12/0.01.01, ABO*Bw.34/A1.02, receptively. Sequencing results showed that there were c.695T>C, c.278C>T, c.889G>A, resulting in variants in Leu232Pro, Pro93Leu and Glu297Lys, receptively. CONCLUSION Bw11, Bw12 and Bw34 subgroups were identified, and gene testing can be used as a supplement to determine the ABO blood group subtypes.
ABO Blood-Group System/genetics* ; Alleles ; Blood Grouping and Crossmatching ; Exons ; Genotype ; Humans ; Phenotype ; Sequence Analysis

OBJECTIVETo explore the molecular mechanism for a boy suspected with 3-methylcrotonyl-CoA carboxylase deficiency by neonatal screening.

METHODSPCR and Sanger sequencing were used to identify potential mutations of MCCC1 and MCCC2 genes. SIFT and Polyphen-2 software was used to predict the effect of variant on the protein function and conservation of the variant across various species. Human Splicing Finder and Swiss-PdbViewer4.1.0 were applied to analyze the possible mechanism of the variant.

RESULTSFor the proband, a compound heterozygous mutation was discovered in the MCCC1 gene, namely c.539G>T (p.G180V) and c.704_711del (p.A235Vfs*4), which were inherited from his father and mother, respectively. The two mutations have disrupted the protein conformation, which in turn may impact the function of MCC protein.

CONCLUSIONThe compound heterozygous mutations of the MCCC1 gene may contribute to the 3-methylcrotonyl-CoA carboxylase deficiency manifested by the patient.

Amino Acid Sequence ; Base Sequence ; Carbon-Carbon Ligases ; chemistry ; deficiency ; genetics ; DNA Mutational Analysis ; Heterozygote ; Humans ; Infant, Newborn ; Male ; Models, Molecular ; Mutation ; Neonatal Screening ; methods ; Protein Conformation ; Sequence Homology, Amino Acid ; Urea Cycle Disorders, Inborn ; diagnosis ; genetics

OBJECTIVETo explore the molecular etiology for a Chinese family affected with isolated methylmalonic acidemia (MMA).

METHODSPotential mutations of MUT, MMAA and MMAB genes in the proband were screened by PCR and Sanger sequencing. The pathogenicity of identified mutations was analyzed using Polyphen2, SIFT, HSF, DNAMAN 6.0 and Swiss-PdbViewer4.1.0 software.

RESULTSTwo novel mutations of the MUT gene, including c.581C>T (p.P194L) and c.1219A>T (p.N407Y), were discovered in the proband, which were inherited respectively from his mother and father. Bioinformatics analysis suggested that both mutations were damaging. The affected codons P194 and N407, both located in the (beta, alpha) 8 barrel domain and to which the substrate methylmalonyl-CoA is bound, are highly conserved across various species. Both mutations can disrupt the space conformation of its protein product, affecting the function of the MCM protein.

CONCLUSIONThe novel mutations of MUT gene probably underlie the isolated MMA in this family.

Adult ; Amino Acid Metabolism, Inborn Errors ; enzymology ; genetics ; Amino Acid Sequence ; Animals ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Female ; Humans ; Infant ; Male ; Methylmalonyl-CoA Mutase ; genetics ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; Pedigree ; Point Mutation ; Sequence Alignment

Objective To investigate the role of neutrophils and their IgA Fc receptor CD89 in the occurrence of Henoch-Sch（o）nlein purpura (HSP),to evaluate their effects on vascular endothelial cell apoptosis,and to explore their mechanisms.Methods Peripheral blood neutrophils were isolated from 30 children with acute HSP and 9 age-matched healthy controls separately.After isolation of serum IgA by Jacalin affinity chromatography,IgA was purified by polypropylene dextran gel chromatography.Real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were performed to determine the mRNA and protein expression of CD89 on neutrophils respectively,and flow cytometry was conducted to measure the expression of neutrophil activation marker CD11b.Human umbilical vein endothelial cells (HUVEC) were co-cultured with neutrophils isolated from patients with HSP (HSP group) and healthy controls (healthy control group) separately.Moreover,the HSP group were divided into 3 subgroups to be treated with serum IgA isolated from the HSP patients (HSP IgA group),monomeric IgA (mIgA group) and phosphate-buffered saline (blank control group) respectively.Then,flow cytometry was conducted to detect apoptosis of co-cultured HUVEC,and enzyme-linked immunosorbent assay (ELISA)to measure levels of interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) in the supernatant of co-cultured cells.Results There was no significant difference in the mRNA expression of CD89 on neutrophils between the patients with HSP and healthy controls (P =0.98),but the protein expression of CD89 was significantly lower in the patients with HSP than in the healthy controls (0.60 ± 0.16 vs.0.83 ± 0.24,P =0.03).The expression of CD1 1b on neutrophils was significantly higher in the patients with HSP than in the healthy controls (1 880.25 ± 388.29 vs.1 109.25 ± 364.25,P ＜ 0.01).The apoptosis rate of co-cultured HUVEC was also significantly higher in the HSP group than in the healthy control group (37.44％ ± 5.49％ vs.17.14％ ± 4.45％,P ＜ 0.01).In addition,the H SP IgA group showed significantly higher apoptosis rate of cocultured HUVEC and levels of IL-8 and TNF-cα in the supematant compared with the mIgA group (all P ＜0.01) and blank control group (P ＜ 0.01,=0.01,=0.02,respectively).Conclusions Peripheral blood neutrophils in patients with HSP are activated,which can induce the apoptosis of vascular endothelial cells.HSP IgA can promote the neutrophil-mediated apoptosis of vascular endothelial cells and secretion of IL-8 and TNF-α,while mIgA may show a certain inhibitory effects.