Genetic diseases that have shown clinical symptoms in neonatal period are often characterized with atypical symptoms and serious condition, which contributes to difficulties in diagnosis and treatment.With the progress of sequencing technology, the next-generation sequencing technology is gradually applied to the clinical field with its advantages of high throughput, low cost and rapid detection.As one of next-generation sequencing technologies, the whole exome sequencing technology(WES)captures, enriches and sequences the genomic exon regions, and then the large amount of WES data is analyzed by bioinformatics methods and screened to find variant site in gene that causes genetic disease.WES technology has gradually become an important means of diagnosis in neonatal genetic diseases because of its advantages of comprehensive results and short reporting period.

ObjectiveTo investigate the diagnostic value of mini peak flow meters in cough variant asthma (CVA). Methods131 patients with the main symptom of chronic cough whose chest X-ray showed normal were included in this study. The peak expiratory flow (PEF) rate of each patient was measured by mini peak flow meters during the bronchial dilation test, and the results were analyzed. ResultsThe rate of positive reaction in the test was 33.6% (44/131), of whom 40.9% (18/44) were delayed over 1 year. 16.8% (22/131) of all the patients' PEF increased 10% to 15% in after the test. ConclusionMini peak flow meters can be used in the diagnosis of CVA.

Objective To study the status of mouse fetal transgenic cells existed in the maternal blood in peri-natal period and provide the information for gene diagnosis using fetal cells circulated in the maternal blood.Methods Mating the female normal mice with male transgenic mice integrated with GFP reporter gene driven by ?-globin promoter and HS2-HS3 elements of LCR. Around the offspring were born, maternal blood was collected and GFP expression level was determined with FACS analysis. Meanwhile, DNA extracted from the tails of the mothers and their offspring as well as the maternal blood were analyzed by PCR.Results GFP positive fetal cells were not found in maternal blood before offspring were born. 1-3 weeks later, GFP positive fetal cells were detected and this population in maternal peripheral blood reached the peak. 4-5 weeks later, they disappeared gradually. PCR results showed no GFP positive band in the mothers. However, a positive fragment of GFP gene was amplified in maternal blood samples after their offspring were born. This result was in accordance with FACS analysis.Conclusion Transgene cells may be useful markers for the study of status of fetal cells existed in the maternal circulating blood. The results would be beneficial for the gene diagnosis using maternal blood as an alternative resource of fetal cells.

Objective: To explore the diagnostic value of ultrasound imaging for fetal central nervous system(CNS) malformation in early pregnancy. Methods: From January 2017 to January 2018, a total of 190 cases of CNS malformation in early pregnancy, who were diagnosed by ultrasound imaging in Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine were selected to analyze the clinical value of ultrasound imaging in the diagnosis of fetal CNS malformation in early pregnancy. Results: Of the 190 pregnancies, 184 patients had satisfactory median sagittal section of the head and chest, horizontal cross section of the lateral ventricle, long axis of the spine, and cerebellum and posterior fossa pool.In 184 cases, the probability of display from 1 to 11^{+ 6} weeks was 93.33%, the probability of display from 12 to 12^{+ 6} weeks was 97.20%, and the probability of display from 13 to 13^{+ 6} weeks was 97.06%.Compared the display rates of CNS ultrasound examinations at different gestational weeks, the difference was not statistically significant (χ²=0.981, P>0.05). Three cases of 6 cases of CNS malformation diagnosed in early pregnancy were combined with other malformations.One case considered Cantrell's hypothyroidism with CNS malformation, 1 case with left iliac crest and right circumflex, 1 case with median cleft lip and palate, single atrioventricular with pulmonary atresia.The bilateral humerus was missing, in addition, 4 fetuses had an NT value more than 3 mm.The specificity of diagnosis of fetal CNS malformation in early pregnancy was 100.00%, the sensitivity was 85.71%, the specificity was 100.00%, and the positive predictive value was 99.46%. Conclusion Ultrasound imaging in the diagnosis of fetal CNS malformation in early pregnancy can improve the diagnosis rate and has high clinical value.

OBJECTIVETo identify the human hematopoietic stem cells from the human/goat xenogeneic model with molecular techniques.

METHODSDNA and total RNA were extracted from 11 transplanted goat peripheral blood cells. Human CD(34), GPA and SRY genes were amplified with PCR in these samples, and CD(34), GPA mRNA transcripts were detected using RT-PCR in 5 and 6 goat peripheral blood cells, respectively. Southern blot analysis was performed in 8 goat DNAs to detect the human specific alpha-satellite sequence. Meanwhile FISH was also performed to detect the human cells in goat blood with a probe of human Y chromosome.

RESULTSHuman CD(34) and GPA genes could be detected with PCR in all the 11 goats, and SRY gene did in 5 goats transplanted with hematopoietic stem cells derived from male human babies. Southern blot showed that human specific alpha-satellite sequence was present in 8 goats. By RT-PCR, human CD(34) mRNA was detected in 5 experimental goats, GPA mRNA was found in the other 6 experimental goats and FISH assay showed that some peripheral blood cells of the human/goat xenogeneic model were positive.

CONCLUSIONExistence of human cells in the recipient goats was identified by molecular detection, which was feasible for the examination of human/goat xenogeneic models.

Animals ; Antigens, CD34 ; genetics ; Blotting, Southern ; Female ; Genes, sry ; genetics ; Glycophorin ; genetics ; Goats ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Polymerase Chain Reaction ; Transplantation Chimera ; genetics ; Transplantation, Heterologous

Objective:To observe the clinical effect of two-dimensional speckle tracking technique in diagnosis and prognosis evaluation of heart failure with left ventricular ejection fraction preservation.Methods:A total of 64 heart failure patients with reduced left ventricular ejection fraction (HFNEF) and 64 healthy volunteers in the Integrated Traditional Chinese and Western Medicine Hospital of Wenzhou were selected from July 2016 to July 2018.Conventional echocardiography and two-dimensional speckle tracking were performed in all subjects.The hemodynamic parameters and strain parameters of two-dimensional speckle tracking were recorded.The differences of conventional echocardiographic parameters and two-dimensional speckle tracking strain parameters in different subjects were analyzed.Results:The E/A (0.79±0.24) and E/E' value (18.93±3.41) in HFNEF patients had statistically significant differences compared with healthy subjects [(1.28±0.35) and (6.84±1.37)] ( t=9.507, 23.294; P=0.000, 0.000). There were no statistically significant differences in HR, LAD, LVD, LVSD, LVPWD, LVEF and LVFS between HFNEF patients [(71.37±5.93)times/min, (35.32±4.37)mm, (47.13±5.01)mm, (9.24±1.76)mm, (9.35±1.82)mm, (63.98±5.37)%, (35.49±4.46)%] and healthy subjects [(70.14±5.28)times/min, (37.64±4.69)mm, (49.26±5.54)mm, (9.39±1.82)mm, (9.48±1.90)mm, (66.35±5.41)%, (34.02±4.17)%] ( t=0.384-1.628; P=0.102-0.564). In HFNEF patients, the anterior septum, anterior wall, posterior wall, inferior wall and posterior wall of apical segment of left ventricle, the inferior wall and posterior wall of papillary muscle and the posterior wall, inferior wall and posterior wall of basal segment, SRs and ROT of left ventricle were significantly lower than those of healthy subjects, while GLS (-13.52±4.18) and GCS (-10.43±3.85) of left ventricle in HFNEF patients were significantly higher than those of healthy subjects [(-19.89±4.61) and (-15.67±4.24)] ( t=8.947~14.063; P=0.000-0.000<0.05). After treatment, the clinical symptoms of HFNEF patients improved significantly in the whole apical segment of left ventricle, inferior and posterior wall of papillary muscle segment, posterior lateral wall, inferior wall, posterior wall SRs and left ventricular GLS, GCS and ROT ( t=7.258-13.193; P=0.000-0.000<0.05). Two-dimensional strain GLS and GC values in HFNEF patients were negatively correlated with E/A values and positively correlated with E/E' value ( r=-0.817, -0.763; P=0.004, 0.012<0.05), while two-dimensional strain ROT results were positively correlated with E/A values and negatively correlated with E/E' values ( r=-0.805, -0.759; P=0.006, 0.014<0.05). Conclusion:Left ventricular systolic function damage has occurred in HFNEF patients.Two-dimensional speckle tracking technique is highly sensitive to the changes of myocardial systolic function for HFNEF patients.It has high clinical value in the diagnosis and prognosis evaluation of HFNEF.Relevant clinical workers should pay attention to it.

Objective: To evaluate the predictive value of cervical size change rate and morphological distribution detected by ultrasound in the second trimester of pregnancy for preterm delivery. Methods: From June 2016 to June 2018, 300 pregnant women who underwent antenatal testing and gave birth in , the Integrated Traditional Chinese and Western Medicine Hospital of Wenzhou were selected in this research.The length, width and shape of cervix were measured by transvaginal color doppler ultrasonography at 14 and 28 weeks respectively.According to whether or not premature delivery occurred, they were divided into premature delivery group and full-term delivery group.The differences of cervical length, cervical inner mouth width, cervical length shortening rate, cervical inner mouth width increasing rate and cervical shape distribution between preterm and full-term pregnant women were compared, and the correlation between the above indicators and the occurrence of premature delivery were analyzed. Results: The length of cervix in the preterm delivery group [(21.41±6.28)mm] was significantly shorter than that in the full-term delivery group at 28 weeks of gestation [(34.17±5.76)mm](t=10.295, P=0.000), and the width of cervical inner mouth in the preterm delivery group [(9.54±2.57) mm] was significantly longer than that in the full-term delivery group [(4.06±0.91)mm] (t=25.416, P=0.000). The shortening rate of cervical length [(39.28±8.65)%] and the increasing rate of cervical width [(149.74±15.09)%] in the preterm pregnant women during the second trimester were significantly higher than those in the full-term pregnant women [(10.15±5.29)% and (16.33±3.84)%] (t=21.471, 39.420; P=0.000, 0.000). The proportion of T-type cervix in the preterm delivery group (77.17%) was significantly higher than that in the full-term delivery group (63.28%) (χ²=10.935, P=0.001). According to the correlation analysis, the length of cervix was negatively correlated with premature birth (r=-0.612, P=0.035), and the width of cervical inner mouth, the shortening rate of cervical length in the second trimester and the increasing rate of cervical inner mouth width in the second trimester were positively correlated with premature birth (r=0.743, 0.665, 0.807; P=0.013, 0.026, 0.004). Conclusion Ultrasound monitoring of cervical size change rate and cervical morphology during the second trimester of pregnancy is helpful to the early prediction of preterm labor, which should be paid attention to clinically.

OBJECTIVE: To study the correlation between DNA methylation patterns and gene expression in Down syndrome (DS). METHODS: Induced pluripotent stem cells (iPSCs) derived from normal controls and DS patients were subjected to whole genome bisulfite sequencing and differentially methylated region (DMR) screening. Statistical analysis for chromosomal and gene element distribution were carried out for DMR. Gene ontology (GO) and enrichment-based cluster analysis were used to explore the molecular function of differentially expressed genes. RESULTS: A total of 1569 DMR were identified in iPSCs derived from DS patients, for which the proportion of hypermethylation in promoter regions was significantly greater than that of the genebody. No DMR enrichment was noted on chromosome 21. Hypermethylation of the promoter and genebody was predicted to be inhibitory for gene expression. Functional clustering revealed the pathways related to neurodevelopmental, stem cell pluripotency and organ size regulation to be significantly correlated with differentially methylated genes. CONCLUSION Extensive and stochastic anomalies of genome-wide DNA methylation has been discovered in iPSCs derived from DS patients, for which the pattern and molecular regulation of methylation were significantly different from those of normal controls. Above findings suggested that DNA methylation pattern may play a vital role in both the pathogenesis of neurodevelopmental disorders and other phenotypic abnormalities during early embryonic development.
DNA Methylation ; Down Syndrome/genetics* ; Female ; Humans ; Induced Pluripotent Stem Cells ; Pregnancy ; Promoter Regions, Genetic ; Whole Genome Sequencing

Objective:To investigate the clinical efficacy of three-dimensional speckle tracking imaging in the dynamic evaluation of left ventricular systolic function in patients with pregnancy-induced hypertension.Methods:50 patients with pregnancy-induced hypertension, who received prenatal examination in Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from July 2019 to June 2020 and finally gave birth, were included in the observation group. An additional 50 healthy pregnant women who concurrently received prenatal examination were included as controls. All participants underwent routine echocardiography and three-dimensional speckle tracking imaging examinations. Parameters related to left ventricular systolic function were recorded. Routine echocardiography parameters and three-dimensional speckle tracking imaging parameters were compared between the observation and control groups.Results:There were no significant differences in routine echocardiography parameters (including heart rate, left ventricular end-diastolic diameter, interventricular septum thickness in end-diastole, left ventricular posterior wall thickness in diastole, and left ventricular ejection fraction) between before treatment, after treatment, and 3 months after delivery in the observation group, and between observation and control groups before treatment (all P > 0.05). The absolute values of left ventricular global longitudinal strain (LVGLS) [(-18.41 ± 2.23)% vs. (-26.03 ± 2.79)%], left ventricular global circumferential strain (LVGCS) [(31.29 ± 3.09)% vs. (37.45 ± 3.68)%], left ventricular global radial strain (LVGRS) [(-19.37 ± 2.19)% vs. (-24.59 ± 2.74)%], and left ventricular global area peak systolic strain (LVGAS) [(-26.61 ± 3.18)% vs. (-39.23 ± 3.96)%] measured before treatment were significantly lower in the observation group than in the control group ( t = 6.31 -14.87, all P < 0.05). The absolute values of LVGLS, LVGCS, LVGRS, and LVGAS in the observation group were significantly higher after treatment [(-24.79 ± 2.68)%, (35.94 ± 3.25)%, (-22.48 ± 2.41)%, (-37.54 ± 3.38)%] and 3 months after delivery [(-25.64 ± 2.72)%, (36.63 ± 3.47)%, (-23.91 ± 2.69)%, (-38.49 ± 3.64)%] than before treatment ( t = 4.08 - 10.59, P < 0.05). There were no significant differences in LVGLS, LVGCS, LVGRS, and LVGAS between observation group and control groups at each time point studied ( t = 0.47 - 1.19, P = 0.182 - 0.652 > 0.05). The absolute svalues of LVGLS, LVGCS, LVGRS and LVGAS in patients with pregnancy-induced hypertension were positively correlated with left ventricular ejection fraction values ( r = 0.638 - 0.775, P = 0.009 - 0.041 < 0.05). Conclusion:Three-dimensional speckle tracking imaging can be used to dynamically evaluate the impairment of left ventricular systolic function in patients with pregnancy-induced hypertension. This technique helps guide early intervention and prognosis evaluation and has a high clinical application value.

Objective:To investigate the value of abdominal ultrasound in the diagnosis of postpartum diastasis of the rectus abdominis muscle (DRA).Methods:A total of 300 pregnant women who received prenatal examination and finally gave birth in Wenzhou Hospital of Integrated traditional Chinese and Western Medicine between October 2018 and October 2020 were included in this study. All of them underwent finger test and high-frequency ultrasound to determine the degree of DRA. The occurrence of DRA was recorded. The efficacy of abdominal high-frequency ultrasound versus finger test in the diagnosis of DRA was analyzed. The high-frequency ultrasound outcomes regarding DRA distance at different positions and at different phases were compared between women with DRA and those without DRA. Results:The overall incidence of DRA among all women included in this study was 57.67% (173/300). The incidence of DRA in multiparae was significantly higher than that in primipara [73.38% (102/139) vs. 44.10% (71/160), χ2 = 26.20, P = 0.001). The incidence of DRA in women subjected to cesarean section was significantly higher than that in women subjected to vaginal delivery [68.52% (74/108) vs. 51.56% (99/192), χ2 = 8.14, P = 0.004). The sensitivity, specificity and accuracy of abdominal ultrasound in the diagnosis of DRA were 99.42%, 98.42% and 99.00%, respectively, which were significantly higher than those of finger test [80.35%,85.04%, 82.33%, χ2 = 34.61, 15.00, 49.23, all P < 0.01]. The DRA distances at different positions measured at 36 weeks of gestation and 2 weeks postpartum were significantly shorter in women with DRA than those without DRA ( t = 5.17-7.46, P < 0.001). Conclusion:Abdominal ultrasound is of high clinical application value in the early diagnosis and rehabilitation treatment of postpartum DRA.