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OBJECTIVE: This study aims to assess the feasibility and effectiveness of color doppler flow imaging (CDFI) technology and the Slow Flow HD imaging technique in identifying fetal pulmonary veins (PVs) in the first trimester (11-13 + 6 weeks), and further explore the factors affecting fetal pulmonary vein identification in early pregnancy. METHODS: Echocardiography and scanning of PVs were performed in 240 normal singleton fetuses in early pregnancy by using CDFI and slow flow HD techniques, to compare the ability of two methods to identify the PVs. Slow Flow HD technology was used to further investigate the difference of PVs identification at different gestational ages [group I (11-11 + 6 weeks), group II (12-12 + 6 weeks), group III (13-13 + 6 weeks)] and with different maternal body mass indices (BMI) (≥ 25 and < 25). In 31 cases of 240 fetuses, transvaginal ultrasonography was added due to maternal habitus or significant retroversion of the uterus, and the difference in PVs identification between transabdominal and transvaginal examination was analyzed. RESULTS: Successful PVs identification rates via CDFI and Slow Flow HD were 32.0% and 88.3%, respectively (p < 0.05). The identification rate of at least one and two pulmonary veins in Slow Flow HD was 88.3% and 76.2%, and all four pulmonary veins in 11.6% (p < 0.05). The identification rate of group I, II and III were 76.4%, 88.9% and 96.0%, respectively. The identification rate was 45.1% in the transabdominal ultrasound group and 83.8% in the transvaginal ultrasound group. The identification rate was 62.5% in the BMI ≥ 25 group and 94.7% in the BMI < 25 group (p < 0.05). CONCLUSIONS: Slow Flow HD can detect PVs in early pregnancy more often than using CDFI. Slow Flow HD is a feasible and effective imaging technique for evaluating PVs in early pregnancy.
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Estudios de Factibilidad , Primer Trimestre del Embarazo , Venas Pulmonares , Ultrasonografía Doppler en Color , Ultrasonografía Prenatal , Humanos , Femenino , Embarazo , Adulto , Ultrasonografía Prenatal/métodos , Venas Pulmonares/diagnóstico por imagen , Edad Gestacional , Índice de Masa CorporalRESUMEN
OBJECTIVE: To explore the genetic basis for a fetus with limb abnormality and cardiac malformation. METHODS: Clinical data of a fetus diagnosed at the Shandong Provincial Maternal and Child Health Care Hospital on April 30th, 2021 was collected. Whole exome sequencing (WES) was carried out, and candidate variant was verified by Sanger sequencing and bioinformatic analysis. X-inactivation analysis was carried out for the female members of its family. RESULTS: The fetus was found to have meningoencephalocele, absence of bilateral radii, cleft lip, abnormal great arteries, and single umbilical artery at the gestational age of 11+ weeks. Sequencing revealed that the fetus has harbored a hemizygous c.1162del (p.Y388Tfs*7) variant of the FANCB gene, which was maternally inherited. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG) and ClinGen, the variant was classified as pathogenic (PVS1+PM2_Supporting+PP4). X-inactivation analysis has revealed complete skewed X-inactivation in the pregnant woman and her mother. CONCLUSION: The hemizygous c.1162del (p.Y388Tfs*7) variant of the FANCB gene probably underlay the multiple malformations in this fetus.
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Anomalías Múltiples , Labio Leporino , Femenino , Humanos , Embarazo , Proteínas del Grupo de Complementación de la Anemia de Fanconi , Feto , Edad Gestacional , MadresRESUMEN
OBJECTIVE: To assess the value of non-invasive prenatal testing (NIPT) for the detection of fetal copy number variations (CNVs) in addition to trisomies 21, 18, and 13. METHODS: A total of 37 306 pregnant women underwent the NIPT test. For those with fetal CNVs indicated by NIPT and accepted invasive prenatal diagnosis, amniotic fluid samples were obtained for chromosomal karyotyping analysis and chromosome microarray analysis (CMA). All cases were followed up. RESULTS: Among the 37 306 cases, 78 (0.209%) were predicted to have fetal CNVs. Among these, 52 pregnant women accepted invasive prenatal diagnosis, and 15 of them (28.85%) obtained a consistent result. Follow up of 26 women who refused invasive prenatal diagnosis have found 2 cases with spontaneous abortion, 2 with induced labor for fetal malformation indicated by ultrasonography, and 1 had multiple malformations and a consistent result by CMA, which yielded an abnormal rate of 19.23%. CONCLUSION: NIPT can signal fetal chromosomal abnormalities through detection of gain and/or loss of fetal DNA copies. Combined chromosomal karyotyping and CMA can increase the detection rate for common chromosomal aneuploidies and CNVs, thereby provide a basis for genetic counseling for the affected families.
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Variaciones en el Número de Copia de ADN , Cariotipificación , Diagnóstico Prenatal , Trisomía , Aneuploidia , Femenino , Humanos , Embarazo , Trisomía/diagnóstico , Trisomía/genéticaRESUMEN
OBJECTIVE: To explore the mechanism and duration of false positive results of non-invasive prenatal testing (NIPT) caused by vanishing twins. METHODS: To detect the variation of cell-free fetal DNA fraction before and after the fetal death and explore its influence on the results of NIPT at different gestational weeks. Prenatal diagnosis was also carried out on amniotic fluid sample derived from the survivor twin. After birth, the two placentas and papyraceous fetus were obtained to ascertain the definitive genetic diagnosis and pathological changes through fluorescence in situ hybridization, fluorescence quantitative PCR and histopathological examination. Eight cases of vanishing twins leading to discordant NIPT results were reviewed for determining the duration of this influence. RESULTS: The vanishing twin has led to immediate flooding of cfDNA into the maternal plasma due to necrotic cytotrophoblasts, which in turn caused increased release of fetal DNA in a short time. However, this did not change the NIPT result for a period of time. The tissue and chorionic villi of perished fetus presented extensive degenerative necrosis. CONCLUSION: The false positive NIPT result caused by vanishing twins may be attributed to continuous release of DNA fragments into the maternal plasma by the fetuses. The influence of the vanished fetuses, which may lead to discordant NIPT results, can last for at least 7-8 weeks but no more than 12-14 weeks during the first and second trimester.
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ADN , Diagnóstico Prenatal , Femenino , Feto , Humanos , Hibridación Fluorescente in Situ , Embarazo , Segundo Trimestre del EmbarazoRESUMEN
OBJECTIVE: To assess the value of chromosomal microarray analysis (CMA) and next-generation sequencing (NGS) for the analysis of abortic tissues. METHODS: A total of 242 samples of spontaneous abortion were collected and tested by CMA or NGS. RESULTS: The detection was successfully in 238 cases (98.35%). In total 143 cases of chromosomal abnormalities were detected, which accounted for 60.08% of all cases. Numerical chromosomal abnormalities were found in 133 cases(93.01%), structural abnormalities were found in 9 cases (6.29%), and uniparental disomy was found in 1 case(0.70%). CONCLUSION: Both CMA and NGS have the advantages of high-throughput, good coverage, high resolution and rapid analysis. They can be used for the detection of the causes of spontaneous abortions. CMA is more useful for the detection of aneuploidies and uniparental disomy, while NGS has advantages in its throughput, capacity in detecting low percentage chimerism and cost, which can provide more options for clinicians.
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Aborto Espontáneo/genética , Aberraciones Cromosómicas , Secuenciación de Nucleótidos de Alto Rendimiento , Análisis por Micromatrices , Femenino , Humanos , EmbarazoRESUMEN
OBJECTIVE: To assess the value of combined chromosome karyotype analysis and multiplex ligation probe amplification (MLPA) assay for the prenatal diagnosis of fetuses with abnormalities detected by ultrasonography. METHODS: With informed consent obtained, 72 pregnant women with ultrasound detected fetal structural abnormalities underwent percutaneous umbilical cord blood sampling. Routine karyotype analysis and MLPA assay were used to detect potential chromosomal deletions and duplications. RESULTS: Five cases were found with an abnormal karyotype. In addition, the MLPA has detected 2 chromosomal microdeletions and 1 microduplication. Together the two methods have yielded a detection rate of 11.11%. CONCLUSION: For fetal abnormalities revealed by ultrasonography, combined karyotype analysis and MLPA assay can provide a better option for its efficiency and simplicity.
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Cromosomas/genética , Feto/anomalías , Adulto , Femenino , Humanos , Cariotipificación/métodos , Ligadura/métodos , Persona de Mediana Edad , Embarazo , Diagnóstico Prenatal/métodos , Adulto JovenRESUMEN
Background: The potential causes of miscarriage are very complex, including genetic, immune, infectious, and endocrine factors. 50%-60% of miscarriages are caused by chromosomal abnormalities. Chromosomal microarray analysis (CMA) is a key tool in this context, capable of detecting not only copy number variations (CNV) but also loss of heterozygosity (LOH). CMA has been used as a tool to investigate the genetic reasons for miscarriage. Methods: In our study, chromosomal microarray analysis (CMA) conducted 1220 miscarriage villous tissues. The results from this technology were used to identify the genetic reasons for miscarriage and evaluated strategies for subsequent pre-pregnancy planning. Results: Here, the abnormality rate of miscarriage was 56.07%(684/1220). The aneuploidy rate accounted for 81.14%(555/684), and was significantly higher in group >35-year-old age. The second most common genetic reason for miscarriage was polyploidy, accounting for 10.09%(69/684). Additionally, we discovered loss of heterozygosity (LOH) in a small percentage of cases, accounting for 2.20%(15/684) reason for miscarriage genetic reasons, due to the advantage of CMA can detect isodisomy (a kind of uniparental disomy). 45 cases (6.58%) with copy number variants, which due to the CMA can detect copy number variations. Conclusion: Our study indicated that miscarriage villous tissues should be performed genetic analysis, seek help from professional genetic counseling.
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The "Fetal Origins of Adult Disease (FOAD)" hypothesis holds that adverse factors during pregnancy can increase the risk of chronic diseases in offspring. Here, we investigated the effects of prenatal hypoxia (PH) on brain structure and function in adult offspring and explored the role of the N6-methyladenosine (m6A) pathway. The results suggest that abnormal cognition in PH offspring may be related to the dysregulation of the m6A pathway, specifically increased levels of YTHDF3 in the hippocampus. YTHDF3 interacts with BTG2 and is involved in the decay of Cbln1 mRNA, leading to the down-regulation of Cbln1 expression. Deficiency of Cbln1 may contribute to abnormal synaptic function, which in turn causes cognitive impairment in PH offspring. This study provides a scientific clues for understanding the mechanisms of impaired cognition in PH offspring and provides a theoretical basis for the treatment of cognitive impairment in offspring exposed to PH.
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Accumulating evidence suggests that prenatal stress (PNS) increases offspring susceptibility to depression, but the underlying mechanisms remain unclear. We constructed a mouse model of prenatal stress by spatially restraining pregnant mice from 09:00-11:00 daily on Days 5-20 of gestation. In this study, western blot analysis, quantitative real-time PCR (qRTâPCR), immunofluorescence, immunoprecipitation, chromatin immunoprecipitation (ChIP), and mifepristone rescue assays were used to investigate alterations in the GR/P300-MKP1 and downstream ERK/CREB/TRKB pathways in the brains of prenatally stressed offspring to determine the pathogenesis of the reduced neurogenesis and depression-like behaviors in offspring induced by PNS. We found that prenatal stress leads to reduced hippocampal neurogenesis and depression-like behavior in offspring. Prenatal stress causes high levels of glucocorticoids to enter the fetus and activate the hypothalamicâpituitaryâadrenal (HPA) axis, resulting in decreased hippocampal glucocorticoid receptor (GR) levels in offspring. Furthermore, the nuclear translocation of GR and P300 (an acetylation modifying enzyme) complex in the hippocampus of PNS offspring increased significantly. This GR/P300 complex upregulates MKP1, which is a negative regulator of the ERK/CREB/TRKB signaling pathway associated with depression. Interestingly, treatment with a GR antagonist (mifepristone, RU486) increased hippocampal GR levels and decreased MKP1 expression, thereby ameliorating abnormal neurogenesis and depression-like behavior in PNS offspring. In conclusion, our study suggested that the regulation of the MKP1 signaling pathway by GR/P300 is involved in depression-like behavior in prenatal stress-exposed offspring and provides new insights and ideas for the fetal hypothesis of mental health.
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Background: To describe the genetic variation of dynein cytoplasmic 2 heavy chain 1 (DYNC2H1) gene in four Chinese families affected with short-rib thoracic dysplasia 3 with or without polydactyly (SRTD3), and to provide evidence for accurate prenatal diagnosis and genetic counseling. Methods: The detailed clinical prenatal sonographic features of four fetuses with SRTD3 were carried out. Trio-whole exome sequencing (WES) and proband-WES sequencing was applied to filtrated causative variants in four families. The causative variants of each family were validated in by Sanger sequencing. Bioinformation analysis was applied to predict the harmfulness of these mutations and perform the protein-protein interaction network and Gene Ontology (GO) analysis. A vitro minigene splicing assay was conducted to assess the influence of the splice site variant. Results: Typical characterization of the four fetuses included short long bones, short ribs, narrow chest, hand and foot posture abnormalities, femur short in diameter and slightly bowing, cardiac abnormalities, and so on. Moreover, eight compound heterozygous variants of DYNC2H1 (NM_001080463.2): c.3842A>C (p.Tyr1281Ser) and c.8833-1G>A, c.8617A>G (p.Met2873Val) and c.7053_7054del (p.Cys2351Ter), c.5984C>T (p.Ala1995Val) and c.10219C>T (p.Arg3407Ter), c.5256del (p.Ala1753GlnfsTer13) and c.9737C>T (p.Thr3246Ile), were identified. Among which, c.10219C>T (p.Arg3407Terp), c.5984C>T (p.Ala1995Val) and c.9737C>T (p.Thr3246Ile) were reported in ClinVar databases, and c.8617A>G (p.Met2873Val), c.10219C>T (p.Arg3407Ter), c.5984C>T (p.Ala1995Val) were found in HGMD databases. Four variants (c.3842A>C (p.Tyr1281Ser), c.8833-1G>A, c.7053_7054del (p.Cys2351Ter) and c.5256del (p.Ala1753GlnfsTer13) were first reported as novel mutations. According to the ACMG guidelines, c.8617A>G (p.Met2873Val), c.7053_7054del (p.Cys2351Ter), c.5984C>T (p.Ala1995Val), c.10219C>T (p.Arg3407Ter) and c.5256del (p.Ala1753GlnfsTer13) were rated as pathogenic or likely pathogenic variants, others variants were predicted to be variants of uncertain significance mutations. The minigene assay results indicated that c.8833-1G>A caused the skipping over exon 56, resulting in exon 56 loss. Conclusion: In our study, we analyzed the genetic mutations in four fetuses with SRTD3 by whole exome sequencing and identified pathogenic variants causing SRTD3. Our results expand the mutation spectrum of DYNC2H1 in SRTD3, which is helpful for the accurate prenatal diagnosis of SRTD3 fetuses and provide useful strategies for genetic counseling.
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Chromosomal structural variations (SVs) are a main cause of human genetic disease. Currently, karyotype, chromosomal microarray analysis (CMA), and fluorescent in situ hybridization (FISH) form the backbone of current routine diagnostics (CRD). These methods have their own limitations. CRD cannot identify cryptic balanced SVs and complex SVs even if these techniques were performed either simultaneously or in a sequential manner. Optical genome mapping (OGM) is a novel technology that can identify several classes of SVs with higher resolution, but studies on the applicability of OGM and its comparison with CRD are inadequate for difficult and complicated chromosomal SVs are lacking. Herein, seven patients with definite complicated SVs involving at least two breakpoints (BPs) were recruited for this study. The results of BPs and SVs from OGM were compared with those from CRD. The results showed that all BPs of five samples and partial BPs of two samples were detected by OGM. The undetected BPs were all close to the repeat-rich gap region. Besides, OGM also detected additional SVs including a cryptic balanced translocation, two additional complex chromosomal rearrangement (CCR). OGM yielded the additional information, such as the orientation of acentric fragments, BP positions, and genes mapped in the BP region for all the cases. The accuracy of additional SVs and BPs detected by OGM was verified by FISH panel and next-generation sequencing and Sanger sequencing. Taken together, OGM exhibit a better performance in detecting chromosomal SVs compared to the CRD. We suggested that OGM method should be utilized in the clinical examination to improve the efficiency and accuracy of genetic disease diagnosis, supplemented by FISH or karyotyping to compensate for the SVs in the repeat-rich gap region if necessary.
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Aberraciones Cromosómicas , Reordenamiento Génico , Humanos , Hibridación Fluorescente in Situ , Mapeo Cromosómico/métodos , CromosomasRESUMEN
Parkinson's disease (PD) is one of the most common degenerative diseases. It is most typically characterized by neuronal death following the accumulation of Lewis inclusions in dopaminergic neurons in the substantia nigra region, with clinical symptoms such as motor retardation, autonomic dysfunction, and dystonia spasms. The exact molecular mechanism of its pathogenesis has not been revealed up to now. And there is a lack of effective treatments for PD, which places a burden on patients, families, and society. CRISPR Cas9 is a powerful technology to modify target genomic sequence with rapid development. More and more scientists utilized this technique to perform research associated neurodegenerative disease including PD. However, the complexity involved makes it urgent to organize and summarize the existing findings to facilitate a clearer understanding. In this review, we described the development of CRISPR Cas9 technology and the latest spin-off gene editing systems. Then we focused on the application of CRISPR Cas9 technology in PD research, summarizing the construction of the novel PD-related medical models including cellular models, small animal models, large mammal models. We also discussed new directions and target molecules related to the use of CRISPR Cas9 for PD treatment from the above models. Finally, we proposed the view about the directions for the development and optimization of the CRISPR Cas9 technology system, and its application to PD and gene therapy in the future. All these results provided a valuable reference and enhanced in understanding for studying PD.
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Amyotrophic lateral sclerosis (ALS) is a deadly neurological disease with a complicated and variable pathophysiology yet to be fully understood. There is currently no effective treatment available to either slow or terminate it. However, recent advances in ALS genomics have linked genes to phenotypes, encouraging the creation of novel therapeutic approaches and giving researchers more tools to create efficient animal models. Genetically engineered rodent models replicating ALS disease pathology have a high predictive value for translational research. This review addresses the history of the evolution of gene editing tools, the most recent ALS disease models, and the application of CRISPR/Cas9 against ALS disease.
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Purpose: To identify potential demographic and lifestyle factors associated with progression of myopia with orthokeratology (ortho-k) treatment via follow-up of axial length (AL). Methods: In this retrospective observational study, demographics, ocular parameters, near-work distance, outdoor activities, and sleep quality were analyzed in 134 children with myopia aged 8~15 years using ortho-k and a follow-up for one year. Results: Compared with the slow progression group, the participants in the fast progression group were younger in age (10.55 ±1.70 years vs 9.90 ±1.18 years, P = 0.009), demonstrated higher spherical equivalent (SE) value (-2.52 ±0.63 diopters (D) vs -3.05 ±0.89 D, P < 0.001), shorter near-work distance (P = 0.010), and poorer sleep quality (Pittsburgh sleep quality index [PSQI], 4.79 ±1.29 vs 3.81 ±1.38, P < 0.001) in the one-year follow-up. Furthermore, multivariate linear regression analyses showed that baseline age (B =-0.020, P = 0.020), SE (B = 0.0517, P < 0.001), and total PSQI (B=0.026, P = 0.001) were associated with axial elongation. Advanced logistic regression analyses demonstrated that shorter average near-work distance (P = 0.034), higher SE value (P = 0.023), and poorer sleep quality (P = 0.003) were associated with fast axial elongation. Conclusion: Sleep quality is one of the key factors associated with axial elongation in children with myopia after using ortho-k for one year. Further studies are required to confirm this observation and expand its practical applications.
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BACKGROUND: Hemophilia A (HA) is the most frequently occurring X-linked bleeding disorder caused by heterogeneous variants in the F8 gene, one of the largest genes known. Conventional molecular analysis of F8 requires a combination of assays, usually including long-range polymerase chain reaction (LR-PCR) or inverse-PCR for inversions, Sanger sequencing or next-generation sequencing for single-nucleotide variants (SNVs) and indels, and multiplex ligation-dependent probe amplification for large deletions or duplications. MATERIALS AND METHODS: This study aimed to develop a LR-PCR and long-read sequencing-based assay termed comprehensive analysis of hemophilia A (CAHEA) for full characterization of F8 variants. The performance of CAHEA was evaluated in 272 samples from 131 HA pedigrees with a wide spectrum of F8 variants by comparing to conventional molecular assays. RESULTS: CAHEA identified F8 variants in all the 131 pedigrees, including 35 intron 22-related gene rearrangements, 3 intron 1 inversion (Inv1), 85 SNVs and indels, 1 large insertion, and 7 large deletions. The accuracy of CAHEA was also confirmed in another set of 14 HA pedigrees. Compared with the conventional methods combined altogether, CAHEA assay demonstrated 100% sensitivity and specificity for identifying various types of F8 variants and had the advantages of directly determining the break regions/points of large inversions, insertions, and deletions, which enabled analyzing the mechanisms of recombination at the junction sites and pathogenicity of the variants. CONCLUSION: CAHEA represents a comprehensive assay toward full characterization of F8 variants including intron 22 and intron 1 inversions, SNVs/indels, and large insertions and deletions, greatly improving the genetic screening and diagnosis for HA.
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Hemofilia A , Humanos , Hemofilia A/diagnóstico , Hemofilia A/genética , Factor VIII/genética , Pruebas Genéticas , Intrones , Reacción en Cadena de la Polimerasa Multiplex , MutaciónRESUMEN
OBJECTIVE: To establish the median of serum markers for second trimester screening in Qingdao region and to assess the influence of median correction on the performance of screening. METHODS: Maternal serum alpha-fetoproteins (AFP), human chorionic gonadotrophin, free beta subunit (ß -HCG) and unconjugated oestriol (uE3) were assayed for prenatal screening of 18 188 singleton pregnancies at 15-20(+ 6) weeks gestation from January 2009 to July 2010. The median of serum markers was calculated based on above results and applied for risk estimation in screening for fetal aneuploidy from August 2010 to March 2011. The screening performance, specified in terms of detection rates (DRs), false positive rates (FPRs) and odds of being affected given a positive result (OAPR) were compared between the two groups. The risks of 45 affected pregnancies detected during the study were estimated with both Caucasian and corrected medians. RESULTS: The average level of AFP in local pregnancies was similar to that of the Caucasian population, whilst ß -HCG and uE3 were respectively 11% and 33% higher than those of Caucasians. The multiple of median (MoM) value was between 0.94 and 1.02 for the dataset based on the corrected median. At a cut-off of l in 270, FPR has decreased from 5.2% to 4.9%, and DR of Down syndrome has increased from 60% to 69.2%, and OAPR has increased from 1:79 to 1:59 when evaluating risk based on the corrected median. For the 45 affected pregnancies, three Down syndrome pregnancies could be missed because their risk estimates were lower than the cut-off level based on Caucasian median. CONCLUSION: It is useful to establish and apply population and laboratory-specific medians in order to improve the performance of prenatal screening and diagnosis.
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Estriol/sangre , Hexaclorociclohexano/sangre , Diagnóstico Prenatal/métodos , alfa-Fetoproteínas/análisis , Adulto , Biomarcadores/sangre , Femenino , Humanos , Embarazo , Segundo Trimestre del EmbarazoRESUMEN
OBJECTIVES: To investigate whether aquaporin 4 (AQP4) gene can affect the activation of glial cells and cause the injury of retina of chronic high intraocular pressure mice models, and to discuss its possible mechanism. METHODS: Experimental study. The chronic high intraocular pressure models were established by burning the scleral venous of the right eye, which as the experiment group, and the left eye without any treatment as the control group. The intraocular pressure (IOP) was measured by rebound tonometer. Selected each of the successful model of chronic high intraocular pressure male AQP4 knockout mice (AQP-/-) and their wild-type (WT) male mice 40, divided the two type of mice into five groups after scleral venous burn according to the time of establishing models (24 h, 3 d, 1 w, 2 w, 4 w after scleral venous burn), 8 mouse in each group. And then producing the paraffin sections of mouse eye. Immunohistochemical staining methods was used to observe the expression of the glial fibrillary acid protein (GFAP) in retina glial cells, and observe the expression of the AQP4 in the retina of the WT mouse. Image was acquired under the fluorescence microscopy. The intraocular pressure was analyzed by t-test. RESULTS: After scleral venous burn (24 h, 3 d, 1 w, 2 w, 4 w), there were significant difference (t = 6.66 - 18.08, all P value < 0.05) in the IOP of the AQP4-/- mice (11.30 ± 1.59, 11.20 ± 1.15, 10.60 ± 1.53, 10.75 ± 1.45, 10.45 ± 1.39) and WT mice (11.50 ± 2.56, 11.25 ± 1.65, 10.75 ± 1.33, 10.60 ± 1.33, 10.40 ± 1.19) between the experimental groups and control groups (6.60 ± 0.94, 6.35 ± 0.99, 6.55 ± 0.94, 6.45 ± 0.99, 6.50 ± 0.94 and 6.60 ± 1.05, 6.50 ± 0.89, 6.40 ± 1.09, 6.30 ± 1.13, 6.50 ± 1.05). Since 24 hours after the scleral venous burn, the expression of GAFP of the two type mice began to increase and reached to peak at 1 week after burning. This peak of WT mice was more obvious than that of AQP4-/- mice. The concentration of GAFP began to decrease at 2 weeks after burning and reached to bottom at 4 weeks later. To the WT mice, the expression of AQP4 was remarkable higher in experimental group than that in control group at 1 week after the scleral venous burn. The expression of AQP4 was related to the expression of GAFP in high intraocular pressure of WT mice at 1 w, 2 w, and 4 w after the scleral venous burn. CONCLUSIONS: The chronic high intraocular pressure models can be established successfully by burning the scleral venous. AQP4 gene can affect the activation of the glial cells in chronic high intraocular pressure mice and lead to the injury of retina.
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Acuaporina 4/genética , Neuroglía/citología , Hipertensión Ocular , Retina/citología , Animales , Proteína Ácida Fibrilar de la Glía/metabolismo , Presión Intraocular , Masculino , Ratones , Ratones Noqueados , Hipertensión Ocular/metabolismo , Hipertensión Ocular/patologíaRESUMEN
Purpose: The exact effects of histone deacetylase 3 (HDAC3) inhibition in DR related retinal ganglion cells (RGCs) death remained unclear. This study is aimed at detecting the influence of HDAC3 on the high-glucose-induced retinal ganglion cell death. Methods: The retinal HDAC3 expression in DR of different time points was analyzed by immunohistochemical assay and western blot. Besides, the expression of HDAC3 and both retinal thickness and RGC loss were analyzed. The effects of HDAC3 inhibitor on cell viability, oxidative stress, and apoptosis in high-glucose- (HG-) treated RGCs were analyzed. Both inflammatory and antioxidative factors were detected by ELISA. Results: Advanced effects of HDAC3 inhibition on the expression of NLRP3 inflammasome were detected using western blots. High HDAC3 expression was detected only in the late DR mice (4 months of diabetes duration) but not early DR mice (2 months of diabetes duration). The immunohistochemical assay showed that HDAC3 expression was correlated with both retinal thickness and RCG contents. HDAC3 inhibitor significantly protected the HG-treated RGCs from damaged cell viability, severe apoptosis, and oxidative stress. Advanced pathway analyses showed that HDAC3 inhibition inactivated NLRP3 inflammasome and thus alleviated retinal inflammation. Conclusion. In conclusion, HDAC3 was involved in RGC loss and thus promoted the progression of neurodegeneration of DR. Besides, HDAC3 inhibitor demonstrated protective effects in neurodegeneration in DR through downregulation of NLRP3 activity. The effects of HDAC3 inhibitor in DR management should be confirmed in clinical trials.
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Inflamasomas , Células Ganglionares de la Retina , Animales , Apoptosis , Glucosa/metabolismo , Inhibidores de Histona Desacetilasas/farmacología , Histona Desacetilasas , Inflamasomas/metabolismo , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Células Ganglionares de la Retina/metabolismoRESUMEN
OBJECTIVE: To provide basis for selecting the suitable method of Down's syndrome biochemical screening in the second trimester pregnancy. METHODS: A total of 30 547 singleton pregnancies between 14 and 20(+ 6) weeks of pregnancy were collected and analyzed for maternal serum alpha-fetoproteins (AFP) and human chorionic gonadotrophin, free beta subunit (beta-HCG) with or without unconjugated estriol (uE3). The screening risks were calculated using the software Lifecycle. The detection rates and the cost of per Down's syndrome detected were calculated and compared. And four different methods were compared in a series of 64 serum samples from Down's syndrome pregnancies. RESULTS: (1) Among the 64 affected cases, the detection rate of Down's syndrome was improved no matter in the double test (DT) or in the triple test (TT) if software Lifecycle (LC) was used to evaluate risks. And it was not suitable to evaluate risks with software 2T-Risks in the triple tests. (2) In the cohort of 30 547 singleton pregnancies, the detection rate of Down's syndrome with project DT-LC, which was double test using AFP and free beta-HCG together with software Lifecycle, and project TT-LC, which was triple test using AFP, free beta-HCG and uE3 together with software Lifecycle, was 56.25% and 57.14%, respectively. The former project was better because it decreased the false positive rate at a lower running cost. CONCLUSION: The DT-LC is an effective screening strategy for second trimester detection of fetal Down's syndrome in mainland China.
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Síndrome de Down/sangre , Síndrome de Down/diagnóstico , Segundo Trimestre del Embarazo/sangre , Diagnóstico Prenatal/métodos , Adulto , Gonadotropina Coriónica Humana de Subunidad beta/sangre , Estriol/sangre , Femenino , Pruebas Genéticas/métodos , Humanos , Embarazo , Diagnóstico Prenatal/economía , Adulto Joven , alfa-Fetoproteínas/metabolismoRESUMEN
Aim: To investigate whether kaempferol exhibits a protective effect on high glucose-induced epithelial-mesenchymal transition (EMT) by mediating the PVT1/miR-214 and PAK4/ß-catenin pathways in SRA01/04 cells. Methods & methods: qRT-PCR and western blot assays were used for gene and protein determination, and migration and invasion assays were conducted. A coimmunoprecipitation assay was used for determining protein interactions. Results: High glucose effectively upregulated PVT1 expression, downregulated miR-214 expression and promoted cell migration and invasion. Kaempferol attenuated high glucose-induced EMT by increasing PVT1 expression and decreasing miR-214 expression. PAK4 was identified as a direct target of miR-214. PAK4 overexpression could rescue the effects of PVT1 deficiency on SRA01/04 cells. Conclusion: Kaempferol ameliorated the regulatory effects of PVT1/miR-214 on high glucose-induced EMT through PAK4/ß-catenin in SRA01/04 cells.