Objective To study basic thyroid stimulating hormone (bTSH) levels impact on outcomes of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in Qinghai.Methods Totally 282 cases with IVF cycles and 93 cases with ICSI cycles were studied prospectively,according to bTSH level,patients were divided into four groups.Reproduction rate,clinical pregnancy rate,miscarriage rate and live birth rate were studied among four groups.Results (1) In 375 cases with IVF/ICSI cycles,bTSH was positively correlated with abortion rate (r=0.42,P=0.04),but live birth rate and growing rate showed negative correlations with bTSH (r=-0.42,-0.28; P=0.04,0.03).bTSH and the number of eggs,the number of fertilized eggs,the number of embryos,biochemical pregnancy rate,and clinical pregnancy rate were no significant correlation (all P＞0.05).(2) Among women at group of ≤1.7,＞1.7 and ≤2.5,＞2.5 and ≤3.5,＞3.5 mU/L,the implantation rates were 28.7％,27.3％,37.7％ and 19.2％,live birth rates were 80.9％,75.0％,82.7％,and 59.8％,abortion rates were 19.0％,15.0％,16.7％,40.1％; they all showed significant difference (all P＜0.05).Abortion rate in women with high bTSH level was higher than that of women with lower bTSH level,however implantation rate,live birth rate in women with high bTSH level were lower.Conclusion When bTSH level is ＞3.5 mU/L,the abortion rate were increased,but live birth rate,rate of implantation were decreased.

Objective To explore the methodology of reconstruction of a three-dimensional model of lid/cheek junction through continuous paraffin sections and stained lid/cheek junction three-dimensional visualization,and to further explore the feasibility and reliability to provide anatomical basis for clinical teaching of plastic and reconstructive surgery.Methods The full size of lid/cheek junction was cut from the specimens,the size of 25 mm× 15 mm× 10 mm,and then embedded in paraffin,and sectioned in thickness of 15 μm for 200 slices;the experimental HE staining and Masson staining were conducted,Sony camera photos using Adobe Photoshop CS 5.1 image processing software for image registration and 3D-Docter software image segmentation were used to give different colors for establishment of complete three-dimensional model.Results Histologically the LOT of the fascia area,the existence of reconstruction and the LOT model were confirmed.It showed the histological characteristics:the Masson stain displayed red,blue and white tissues in color;fascia tissue staining infered intertexture of elastic fibers and collagen fibers in LOT.LOT was the bottom edge of the triangle toward the orbital base with length of 26 mm,31 mm in high,0.8 mm in thick,and area of approximately 4.03 cm2 in size.Layers of skin,orbicularis oculi muscle,orbicularis retaining ligament,middle temporal fascia,periosteum and LOT were visible in the 3D model.By the three-dimensional model of lid/cheek junction,adjacent relationship could be rotated to any angle.Conclusions This initial establishment of a three-dimensional model of the lid/cheek junction confirms that the histological characteristics of lid/cheek junction and the feasibility of the fine structure of soft tissue within the three-dimensional model can be used as a new method for further research.

Objective To investigate the risk factors for acute radiation esophagitis andpneumonitis after radiation therapy in esophageal cancer (EC) patients with diabetes or hypertension.Methods A total of 373 EC patients receiving three dimensional conformal radiation therapy (3D-CRT) or intensity modulated radiation therapy (IMRT) were included in this study.Among these patients,42 showed concurrent with diabetes and 99 with hypertension.Radiation esophagitis or pneumonitis in patients with or without diabetes,and with or without hypertension were monitored in the 1-year follow up,respectively.Results The prevalence of grade 1,2,3 and 4 radiation esophagitis in diabetes and non-diabetes patients was 40.5％,38.1％,14.3％,4.8％ and 66.2％,27.8％,2.7％,1.8％,respectively,while that of the grade 1,2 and 3 radiation pneumonitis in diabetes and non-diabetes patients was 31.0％,16.7％,9.5％ and 30.8％,15.7％,1.2％,respectively.The prevalence of grade 3 or above radiation esophagitis and pneumonitis in patients with diabetes and was significantly higher than those with non-diabetes (x2 =13.573,12.279,P ＜ 0.05).The prevalence of grade 1,2,3 and 4 radiation esophagitis in hypertension and non-hypertension patients was 49.5％,38.4％,8.1％,3.0％ and 68.2％,25.5％,2.6％,1.8％,respectively,while that of the grade 1,2 and 3 radiation pneumonitis in hypertension and non-hypertension patients were 30.3％,18.2％,5.1％ and 31.0％,15.0％,1.1％,respectively.The prevalence of grade 3 or above radiation esophagitis and pneumonitis in patients with hypertension was significantly higher than those with non-hypertension (x2 =5.695、5.422,P ＜ 0.05).Diabetes is an independent risk factor for grade 3 or above acute radiation esophagitis and pneumonitis.Conclusions Diabetes or hypertension might be risk factors for severe radiation esophagitis and pneumonitis in EC patients receiving radiation therapy.

Objective: To explore whether human adipose-derived stem cells (ADSCs) express PD-L1 and Gal-9 and its potential influence factors. Methods: ADSCs isolated from 28 healthy female donors who underwent liposuction of the abdomen or breast tissue were cultured and characterized. The expression of PD-L1 and Gal-9 were detected using flow cytometry. The impact of donor age, body mass index (BMI), donor site and interferon-γ (IFN-γ) on the expression of PD-L1 and Gal-9 was analyzed using multivariate linear regression analysis. Results: The cultured cells fulfilled the criteria for defining MSCs according to the international standards and expressed PD-L1 and Gal-9. Breast-derived ADSCs had higher expression levels of PD-L1 (37.24%±8.20%) and Gal-9 ( 4.41%±2.65%) than those in abdomen-derived ADSCs (28.80%±8.59% and 2.51%±1.39%, respectively) (P=0.018, P=0.039). Multivariate linear regression analysis showed that donor age and site were independent risk factors affecting PD-L1 expression (P=0.009, P=0.006). The expression of PD-L1 decreased by 0.385% for every one year of age increase, and it was 8.58% higher in the breast-derived ADSCs than in the abdomen-derived ADSCs. Donor site was an independent risk factor for Gal-9 expression (P=0.041). Gal-9 positive expression rate in the breast-derived ADSCs was 1.898% higher than that in the abdomen-derived ADSCs. BMI was not a risk factor affecting PD-L1 or Gal-9 expression on ADSCs. The expression of PD-L1 and Gal-9 on ADSCs were significantly upregulated after 48 hours stimulation with 20 ng/ml IFN-γ in vitro. Conclusions Human ADSCs expressed PD-L1 and Gal-9. Donor age, site and IFN-γ treatment were independent risk factors affecting the expression of PD-L1 and Gal-9 on ADSCs.

OBJECTIVE: To carry out genetic testing for two families affected with cobalamin C (cblC) and establish a rapid method for the detection of a hotspot pathogenic variant c.609G>A of the MMACHC gene by using a PCR-high-resolution melting curve (PCR-HRM) method. METHODS: Genomic DNA was extracted from peripheral blood samples of the probands and their parents. Potential variants of the MMACHC gene was analyzed by Sanger sequencing. The c.609G>A variant of the MMACHC gene was screened among 100 healthy children with the PCR-HRM method. RESULTS: Sanger sequencing revealed that proband 1 carried compound heterozygous variants c.394C>T and c.609G>A of the MMACHC gene, while proband 2 carried compound heterozygous variants c.482G>A and c.609G>A of the same gene. PCR-HRM analysis of the two probands and the 100 healthy children were consistent with the Sanger sequencing. CONCLUSION c.609G>A is a hotspot pathogenic variant of the MMACHC gene. The diagnosis of cblC may be rapidly attained through detection by PCR-HRM.

OBJECTIVE: To analyze the clinical and molecular characteristics of a child with very long chain acyl-CoA dehydrogenase deficiency (VLCADD). METHODS: Peripheral blood sample of the patient was collected for the extraction of genomic DNA. Next generation sequencing (NGS) was carried out for the proband. Suspected mutations were validated by Sanger sequencing. RESULTS: The patient, a 12-month-old girl, was admitted for diarrhea, vomiting, fever, poor spirit and decreased blood pressure. During the course of the disease, she also manifested hypertrophic cardiomyopathy, cardiogenic shock, elevated myocardial enzyme kinase, fever and metabolic acidosis, and had died after three days due to ventricular tachycardia and respiratory failure. Genetic testing showed that she has carried heterozygous mutations of of the ACADVL gene, namely c.664G>A (exon 8) and c.1056_1057del (exon 10). Blood screening for metabolic genetic diseases showed increased C12, C14, C16, C18, C14:1, C14:2, C16:1, C4/C3 and C8/C3, accompanied with decreased C0, C0/C16 and C8/C10. VLCADD and secondary carnitine deficiency could not be excluded, which was in keeping with the result of genetic testing. CONCLUSION The child was diagnosed with VLCADD, which may be attributed to the compound heterozygous c.664G>A and c.1056_1057del variants of the ACADVL gene.

OBJECTIVE: To explore the genetic basis for a child with succinate semialdehyde dehydrogenase deficiency. METHODS: Peripheral blood samples of the proband and his parents were collected and subjected to Sanger sequencing. High-throughput sequencing was used to verify the gene variants. Bioinformatic software was used to analyze the pathogenicity of the variant sites. RESULTS: Sanger sequencing showed that the proband carried a homozygous c.1529C>T (p.S510F) variant of the ALDH5A1 gene, for which his mother was a carrier. The same variant was not detected in his father. However, high-throughput sequencing revealed that the child and his father both had a deletion of ALDH5A1 gene fragment (chr6: 24 403 265-24 566 986). CONCLUSION The c.1529C>T variant of the ALDH5A1 gene and deletion of ALDH5A1 gene fragment probably underlay the disease in the child. High-throughput sequencing can detect site variation as well as deletion of gene fragment, which has enabled genetic diagnosis and counseling for the family.
Amino Acid Metabolism, Inborn Errors/genetics* ; Child ; Developmental Disabilities ; Humans ; Infant ; Mutation ; Succinate-Semialdehyde Dehydrogenase/genetics*

Objective To investigate the spectrum of CYP21A2 gene mutation and the correlation between genotype and phenotype in patients with 21-hydroxylase deficiency in Tianjin and surrounding areas.Methods Genomic DNA was extracted from the peripheral blood samples of the proband.Locus-specific PCR,direct sequencing of PCR amplification products,and multiplex ligation-dependent probe amplification were applied to detect pathogenic gene CYP21A2 and the relationship between genotypes and phenotypes was analyzed.Results (1) Of 35 patients with 21-hydroxylase deficiency,25 were classified as salt-wasting phenotype and 10 were simple virilizing phenotype.(2) 69 mutant alleles were detected in a total of 70 alleles in 35 patients.Only one mutant allele was detected in one patient.Two mutant alleles were detected in all other patients,with the mutation detection rate 98.6％.(3) A total of 6 types of mutations were detected,of which c.293-13C/A＞G (I2G) was the most common,accounting for 57.1％ (40/70),followed by 18.6％ (13/70) for large gene deletion or conversion,and 14.3％ (10/70) for p.I173N.In addition,a novel mutation,c.949C＞T (p.R317X),which has not been reported previously,was detected as a pathogenic mutation.(4) Correlation analysis of genotype and phenotype in 35 children showed that the phenotype predicted by genotype was consistent with the actual salt-wasting phenotype in 31 children,and those in three children were inconsistent with the actual clinical phenotype.Conclusion The mutation characteristics of CYP21A2 gene in patients with 21-hydroxylase deficiency in Tianjin and surrounding areas are slightly different from those reported in other regions in China.A mutation c.949C＞T has not been reported,which enriches the mutation spectrum of CYP21A2 gene and provide the foundation for genetic counseling and prenatal diagnosis.

OBJECTIVE: To analyze the molecular etiology of a Chinese child affected with dihydropyrimidinase deficiency. METHODS: Genomic DNA was extracted from peripheral blood samples of the family members. Pathogenic variant was determined by whole exome sequencing and verified by Sanger sequencing. RESULTS: The child was found to harbor homozygous c.905G>A (p.Arg302Gln) variants in exon 5 of the DPYS gene, for which her parents were both heterozygous carriers. CONCLUSION The homozygous c.905G>A (p.Arg302Gln) variants of the DPYS gene probably underlies the dihydropyrimidinase deficiency in the child. Above result has enabled genetic counseling and prenatal diagnosis for this family.
Amidohydrolases/genetics* ; Asian Continental Ancestry Group/genetics* ; Child ; Exons ; Female ; Humans ; Metabolism, Inborn Errors/genetics* ; Mutation ; Pedigree

OBJECTIVE: To explore the genetic basis for a child with 46,XY disorders of sex development (DSD) and explore its genotype-phenotype correlation. METHODS: The child was subjected to whole exome sequencing (WES), and exons 1 to 7 of NR5A1 were subjected to multiplex ligation-dependent probe amplification (MLPA) analysis. RESULTS: The patient presented with rudimentary vulva of a female with Tanner stage 1. B-mode ultrasonography has detected ovary and uterus. The child was found to have a chromosome karyotype of 46,XY. WES revealed that the patient has harbored heterozygous deletion of exon 5 of the NR5A1 gene, which was a novel pathogenic variant inherited from the mother. No abnormality was found in the father. CONCLUSION The main symptoms of 46,XY DSD children are insufficient external genitalia masculinization, for which variants of the NR5A1 gene are an important cause. WES has improved the detection rate of genetic variants and provided a solid basis for genetic counseling of the affected families.
Child ; Disorder of Sex Development, 46,XY/genetics* ; Disorders of Sex Development/genetics* ; Exons/genetics* ; Female ; Genetic Testing ; Heterozygote ; Humans ; Mutation ; Steroidogenic Factor 1/genetics*