1.Application of next-generation sequencing technique in genetic analysis of spontaneous abortion
Yilin GUO ; Maosheng GU ; Li WANG ; Suzhen QU ; Shuwen XUE ; Haijian WANG ; Zhaoling XUAN ; Xiangdong KONG
Chinese Journal of Perinatal Medicine 2018;21(12):808-816
Objective To investigate the value of next-generation sequencing (NGS) technique for genetic analysis of spontaneous abortion. Methods From January to June 2017, 154 patients who visited the First Affiliated Hospital of Zhengzhou University for spontaneous abortion were enrolled. All abortion tissue samples were analyzed by both NGS combined with short tandem repeat (STR) and single nucleotide polymorphism array (SNP-array). Results of the two methods were compared by Chi-square or Fisher's exact test. Results (1) Chromosomal abnormalities were detected in 109 of the 154 cases (70.7%), including 52 (47.7%) of numerical chromosomal abnormalities, 49 (45.0%) of structural chromosomal abnormalities, six (5.5%) of mosaicism, and two (1.8%) of uniparental disomy (UPD). In those 52 cases of numerical chromosome abnormalities, there were 45 of chromosome aneuploidy and seven of polyploidy. The top three numerical chromosomal abnormalities were 45,X (27.0%, 14/52), trisomy 22 (9.6%, 5/52) and trisomy 16 (7.7%, 4/52). Forty-nine structural abnormality cases carried 67 copy number variations (CNV), including 13 pathogenic CNV (pCNV, 19.4%), 24 variants of unknown clinical significance (35.8%) and 30 benign CNV (44.8%). In those 13 pCNVs, two were responsible for microdeletion and microduplication syndromes. (2) SNP-array was successful in 152 cases, but failed in two (1.3%) due to genomic DNA <200 ng. However, NGS technology was successful in all 154 cases and identified chromosomal abnormalities in the two cases that SNP-array had failed. No statistically significant difference was shown in the detection rate of chromosomal abnormalities between SNP-array and NGS technology [70.4% (107/152) vs 67.5% (104/154), χ2=0.293, P=0.588]. (3) No significant difference in the detection of chromosome aneuploidy (six cases in each group, 3.9% vs 3.9%) and mosaicism (45 cases in each group, 29.2% vs 29.6%) was found between NGS technology and SNP-array. Three cases of polyploidy (69, XXX) and two of UPD were identified by SNP-array, but not by NGS. When combined with STR, NGS was able to detect all three cases of polyploidy (69, XXX). (4) Forty-seven structural abnormality cases detected by SNP-array carried 53 CNVs, and 49 detected by NGS carried 67 CNVs. (5) NGS detected ten, three and one more CNVs than SNP-array did when the genome lengths were 100-<500, 500-<1 000 and ≥1 000 kb, respectively. Conclusions NGS can be used to detect chromosomal aneuploidy and mosaicism that can be identified by SNP-array with fewer limitations on total amount of genome. Moreover, CNVs that fail to be identified by SNP-array can also be detected by NGS. When combined with STR, NGS can effectively detect chromosomal polyploidy. Therefore, NGS could be a potential genetic analysis method for spontaneous abortion and of importance for genetic counseling.
2.Study on the normal value of cervical elasticity parameters of primiparas during pregnancy by transvaginal ultrasound
Yuan GAO ; Jianjun YUAN ; Yujuan GUO ; Zhaoling ZHU ; Xijun ZHANG ; Li WANG ; Lina WU ; Ruili WANG
Chinese Journal of Ultrasonography 2022;31(8):724-729
Objective:To investigate the normal parameter values of transvaginal E-Cervix cervical elastography of primipara and explore the correlation between E-Cervix parameters and gestational age.Methods:A total of 301 primiparas underwent E-Cervix by transvaginal ultrasound in Henan Provincial People′s Hospital from March 2019 to January 2021, they were divided into 6 groups according to generational weeks: group A (11-13 + 6W), group B (14-17 + 6W), group C (18-21 + 6W), group D (22-25 + 6W), group E (26-29 + 6W), group F (30-33 + 6W). Real-time E-Cervix parameters were obtained, including elasticity contrast index (ECI), hardness ratio (HR), internal ostium (IOS), and external ostium (EOS), strain ratio of the internal and external cervical ostium (IOS/EOS), and the correlations between E-Cervix parameters and the gestational week were analyzed. Results:The normal reference ranges of parameters ECI, IOS, EOS, HR and IOS/EOS were obtained, their 95% reference values in groups A to F were as follows: ECI (1.84-3.38, 2.13-4.50, 2.25-4.12, 2.06-3.96, 2.04-5.15, 2.36-5.53), HR (85.85%-88.72%, 77.31%-80.92%, 74.41%-78.50%, 71.13%-74.35%, 64.49%-68.56%, 63.57%-68.23%), IOS (0.12-0.21, 0.14-0.34, 0.14-0.31, 0.19-0.40, 0.19-0.56, 0.21-0.49), EOS (0.12-0.30, 0.14-0.34, 0.16-0.41, 0.20-0.39, 0.22-0.58, 0.22-0.54), IOS/EOS (0.55-1.12, 0.52-1.45, 0.56-1.26, 0.54-1.38, 0.54-1.52, 0.65-1.46). The differences of ECI, HR, IOS and EOS in different groups were statistically significant (all P<0.05). Spearman correlation analysis results showed that ECI, IOS, and EOS were positively correlated with gestational week( rs=0.524, 0.689, 0.609; all P<0.01). HR was negatively correlated with gestational week ( rs=-0.755, P<0.01). The regression equations of ECI, HR, IOS and EOS were Y=0.06X+ 52.214, Y=99.25-1.07X, Y=8.63E-3X+ 0.05, Y=7.94E-3X+ 0.1, respectively. Conclusions:The normal parameters values of E-TVES are summarized.It provides effective support in data for cervical evaluation in the preterm prediction and success of labor induction.
3.Ultrasonic evaluation of fetal cerebral sulci and gyrus development in pregnant women with gestational diabetes mellitus
Xiaolin ZHANG ; Zhaoling ZHU ; Ruili WANG ; Yuan GAO ; Bingbing LIU ; Liangjie GUO ; Jianjun YUAN ; Jingge ZHAO
Chinese Journal of Ultrasonography 2024;33(1):36-41
Objective:To evaluate the development of fetal cerebral sulci and gyrus and the blood perfusion in pregnant women with gestational diabetes mellitus(GDM) by ultrasound.Methods:A total of 1 540 pregnant women with 28-34 weeks of pregnancy who underwent systematic screening in Henan Provincial People′s Hospital from January 2022 to October 2022 were prospectively selected, 100 pregnant women with GDM were selected as the GDM group. According to the effect of blood glucose control, the GDM group was divided into 2 groups: the satisfied control group (GDM group 1), and the dissatisfied control group (GDM group 2), with 50 cases in each group. At the same period, 50 healthy pregnant women at 28-34 weeks of gestation were enrolled as the control group. The differences of the sylvian fissure, parietooccipital sulci, calcarine sulci and cinguli sulci among the 3 groups were statistically analyzed. And the correlations between the deep of the brain cerebral sulci and gyrus and controlled blood glucose levels were evaluated. The umbilical artery pulsation index(UAPI), middle cerebral artery pulsation index(MCAPI) and ductus venosus pulsation index(DVPI) among the 3 groups were compared, and the differences in fetal blood perfusion among the 3 groups were evaluated.Results:There were no significant differences in the depths of the sylvian fissure, parietooccipital sulci, calcarine sulci and cinguli sulci between the control group and the GDM group 1 (all P>0.05), and they were larger than those of the GDM group 2 (all P<0.05). The depths of lateral fissure, parieto-occipital sulcus, cingulate sulcus and calcarine sulcus were negatively correlated with fasting blood glucose, 1 h and 2 h postprandial blood glucose (all P<0.05). There were no significant differences in MCAPI, UAPI and DVPI between the control group and GDM1 group (all P>0.05). The MCAPI in GDM 2 group was lower than that in the control group and GDM 1 group, and the UAPI and DVPI values were higher than those in the control group and GDM1 group(all P<0.05). Conclusions:The maturity of fetal cerebral sulci and gyrus in GDM pregnant women is related to the blood glucose control of pregnant women. The change of blood perfusion caused by persistent hyperglycemia in pregnant women and intrauterine hypoxia may cause the development retardation of cerebral sulci and gyrus.
4.Analysis of related factors for clinical characteristics and the outcome in centenarian hospitalized patients
Yu WANG ; Weiwei SONG ; Xiaoli CHEN ; Zhiyong WANG ; Jian DAI ; Xiaojun OUYANG ; Lili LIU ; Yu LIU ; Peng ZHANG ; Zhaoling GUO ; Yunyan WEI ; Jihai CHEN ; Weiwei YUAN ; Weihong ZHAO ; Jianqing WU ; Wei XU
Chinese Journal of Geriatrics 2019;38(1):4-9
Objective To investigate the health status of centenarian hospitalized patients and analyze the risk factors for in-hospital death in Nanjing district.Methods All centenarians hospitalized patients who were discharged from wards of 10 upper first-class general hospitals in Nanjing district during the past five years were retrieved from their hospital information systems.Then,a retrospective study was performed on centenarians' data of general information,laboratory test results,Charlson comorbidity index (CCI),neutrophil to lymphocyte ratio (NLR) and shock index(SI),etc.were calculated and collected.Relevant risk factors for in-hospital death were analyzed by multivariate logistic regression analysis.Results A total of 156 patients aged 100 years and over,with an average age of (101.0±2.1)years,were enrolled during the past 5 years.The top 3 admitting diagnosis for the patients were pulmonary infection(30.1%,47/156 cases),coronary heart disease(10.9%,17/156 cases)and cerebrovascular disease(7.1%,11/156 cases).Fifty patients died during hospitalization,with a mortality of 32.1% (50/156).Pneumonia was the most common admitting diagnosis(40.0%,20/50 case).Among causes of death,the combined admitting diagnosis with dementia,chronic renal insufficiency,one or more basic disease were significantly associated with death.There were statistically significant differences between bad vs.good vs.indifferent prognosis in heart rate,shock index,leukocyte count,neutrophil count,NLR,hemoglobin,albumin,albumin/globulin,fasting blood glucose,blood urea nitrogen,serum creatinine,C-reactive protein(CRP)and CCI levels.Multivariate logistic regression analysis suggested that NLR≥13.18,fasting blood glucose ≥7.56 mmol/L,blood urea nitrogen ≥20.74 mmol/L,CRP≥65 mg/L and CCI≥3 might be predictors for in-hospital death in the cohort(OR =48.91、3.43、1.22、6.55、1.55,all P<0.05).Conclusions Pulmonary infection is the most common reason for admission and the cause of death in centenarian inpatients.Comorbidities increase the risk of death.To lower in-hospital mortality,CCI and other assessment indicators should be used to strengthen the comprehensive assessment and chronic disease management of hospitalized centenarians.Infectious diseases should be prevented beforehand.
5.Research Progress on Medical Imaging and New Ultrasound Techniques for Assessing the Degree of Carotid Artery Stenosis
Yigang DU ; Shengli WANG ; Zhaoling LU ; Yanbo LIU ; Yuexin GUO ; Xing AN ; Shuangshuang LI ; Lei ZHU
Chinese Journal of Medical Instrumentation 2024;48(6):624-630
The paper summarizes the imaging evaluation methods for assessing the degree of carotid artery stenosis and analyzes the unique advantages and limitations of various imaging techniques in vascular imaging based on existing guidelines and consensus.The paper focuses on reviewing the clinical applications of several novel ultrasound technologies,including the use of advanced hemodynamic parameters such as blood flow dispersion(Tur index)and wall shear stress(WSS).Carotid artery stenosis is closely associated with cardiovascular disease.Although non-invasive and radiation-free ultrasound technology has certain limitations in diagnostic accuracy to a certain extent,with the continuous emergence of advanced functions such as ultrasound hemodynamics and vascular elasticity,the combination of multi-modality and multi-parameter ultrasound is expected to become an important method for efficient diagnosis of arterial stenosis in the future.
6.Experimental study to construct the three-dimensional scaffold of rabbit cartilage-shedding cells and evaluate their compatibility with stem cells
Juju XU ; Yanyan XIE ; Zhiyi GUO ; Yukai MA ; Linhong WANG ; Meirong YANG ; Zhaoling DENG ; Baolai HUA ; Zhenyu YAN
Acta Laboratorium Animalis Scientia Sinica 2024;32(8):1012-1022
Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells to provide an experimental basis for cartilage repair.Methods Bone marrow mesenchymal stem cells(BMSCs)were cultured using the Percoll density gradient separation method,and this was followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints and subjected to physical crushing,repeated freeze-thaw cycles,and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of the decellularized scaffolds at different concentrations,three groups of scaffolds(labelwd A,B,and C)were designed with concentrations of 100%,50%and 30%,with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto optimally concentrated scaffolds and cultured for 1 week to observe cell growth.Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45.Osteogenic induction stained with alizarin red showed red calcific nodules,and chondrogenic induction stained with alcian blue showed blue cartilaginous nodules.No apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin-eosin,and toluidine blue.There was a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds(P<0.05).The content of glycosaminoglycans was slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size,water absorption,porosity,tensile strength,and Young's modulus(P<0.05).After co-cultivation of stem cells with the scaffolds,cell adhesion was found to be good.Conclusions Percoll density gradient separation can obtain high-purity rabbit BMSCs,and the mixed decellularization method is superior.Group B scaffolds were the most suitable for tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group B scaffolds.