ABSTRACT
BACKGROUND: Evidence from the Istanbul consensus workshop suggests correlations between morphological parameters and embryo developments. 8-cell embryos are the best blastomere stage on day 3. No good quality evidence exists to support high-quality embryonic selection following blastulation and clinical outcomes. This study aimed to investigate the factors that affect blastocyst formation, blastocyst quality, and clinical outcomes of high-quality cleavage-stage embryos in fresh cycles. METHODS: This study was a retrospective analysis of 9608 high-quality cleavage-stage embryos from 2987 couples between January 2017 to June 2021, namely 1520 embryos categorized as "812" (8-cell, grade 2, mild fragmentation), 2961 as "821" (8-cell, grade 2, mild asymmetry), 896 as "711" (7-cell, grade 1), and 517 as "911" (9-cell, grade 1) compared with 3714 embryos categorized as "811" (8-cell, grade 1). The primary outcomes were clinical pregnancy rate (CPR) and live birth rate (LBR). Blastulation rate (BR), available late blastocyst rate (ABR) and high-quality late blastocyst rate (HBR) were secondary outcome measures. RESULTS: BR, ABR, and HBR had significant differences among the five groups (P < 0.001), while CPR and LBR were also significantly different in cleavage-stage fresh transfer (P < 0.01). The multivariable multilevel logistic regression analysis revealed a significant association between cell number, cell size, blastocyst development and clinical outcomes. For 7 to 9-cell highest-quality embryo, mild fragmentation and more blastomeres were more conducive to blastocyst formation and clinical outcomes. While cleavage-stage embryos developed into blastocysts, the negative impact of their initial morphology on clinical outcomes would be erased. CONCLUSIONS: Our study firstly evaluated blastocyst development and clinical outcomes of high-quality cleavage-stage embryos in fresh cycles, with rankings of 811, 812, 911, 821, and 711. We found the initial morphological characteristics of the high-quality cleavage-stage embryos did not adversely impact clinical outcomes, even as they progressed to the blastocyst stage.
Subject(s)
Birth Rate , Embryo Transfer , Pregnancy , Female , Humans , Retrospective Studies , Pregnancy Rate , Embryonic Development , Blastocyst , Live BirthABSTRACT
BACKGROUND: To explore the relationships between hyperuricemia and the risk of cardiovascular diseases (CVD) and chronic kidney disease (CKD) in both the general population and hypertensive patients through meta-analysis. METHODS AND RESULTS: We systematically searched PubMed, Embase, and Cochrane Library databases from January 2012. The eligibility criteria were predefined, and quality was assessed using the Newcastle-Ottawa Scale (NOS). Stata 15.1 was used for meta-analysis, heterogeneity and sensitivity analysis. Subgroup analysis was used to explore heterogeneity, funnel plots and Egger tests were used to assesse publication bias and applicability. A total of 10,662 studies were retrieved, 45 of which were included in this meta-analysis utilizing a random effects model. Hyperuricemia was significantly associated with an increased risk of new-onset hypertension (RR = 1.36, 95% CI 1.16-1.59; I2 = 98.8%), total CVD (RR = 1.53, 95% CI 1.23-1.89; I2 = 93.7%), stroke (RR = 1.97, 95% CI 1.71-2.26, I2 = 0.0%), coronary heart disease (CHD) (RR = 1.56, 95% CI 1.06-2.30, I2 = 93.3%), and CKD (RR = 1.71, 95% CI 1.56-1.87; I2 = 87.3%). However, subgroup analysis showed no significant associations between hyperuricemia and hypertension in non-Asian populations (RR = 0.88, 95% CI 0.59-1.33), or between hyperuricemia and CVD with a follow-up duration <5 years (RR = 1.26, 95% CI 0.97-1.63). Among hypertensive patients, hyperuricemia was significantly associated with total CVD (RR = 2.32, 95% CI 1.31-4.12, I2 = 90.2%), but not with stroke (RR = 1.48, 95% CI 0.86-2.55; I2 = 90.7%) or CHD (RR = 1.51, 95% CI 0.98-2.33; I2 = 71.7%). CONCLUSION: Hyperuricemia was significantly associated with an increased risk of new-onset hypertension, total CVD, stroke, CHD, and CKD in the general population. Among hypertensive patients, hyperuricemia was associated with an increased risk of CVD but not stroke or CHD alone. REGISTRATION NUMBER: CRD42022370692.
Subject(s)
Cardiovascular Diseases , Hypertension , Hyperuricemia , Renal Insufficiency, Chronic , Hyperuricemia/epidemiology , Hyperuricemia/complications , Hyperuricemia/blood , Humans , Renal Insufficiency, Chronic/epidemiology , Renal Insufficiency, Chronic/complications , Hypertension/epidemiology , Hypertension/complications , Cardiovascular Diseases/epidemiology , Risk FactorsABSTRACT
BACKGROUND: The validation of various risk scores in elderly patients with comorbid atrial fibrillation (AF) and acute coronary syndrome (ACS) has not been reported. The present study compared the predictive performance of existing risk scores in these patients. METHODS: A total of 1252 elderly patients with AF and ACS comorbidities (≥ 65 years old) were consecutively enrolled from January 2015 to December 2019. All patients were followed up for one year. The predictive performance of risk scores in predicting bleeding and thromboembolic events was calculated and compared. RESULTS: During the 1-year follow-up, 183 (14.6%) patients had thromboembolic events, 198 (15.8%) patients had BARC class ≥ 2 bleeding events, and 61 (4.9%) patients had BARC class ≥ 3 bleeding events. For the BARC class ≥ 3 bleeding events, discrimination of the existing risk scores was low to moderate, PRECISE-DAPT (C-statistic: 0.638, 95% CI: 0.611-0.665), ATRIA (C-statistic: 0.615, 95% CI: 0.587-0.642), PARIS-MB (C-statistic: 0.612, 95% CI: 0.584-0.639), HAS-BLED (C-statistic: 0.597, 95% CI: 0.569-0.624) and CRUSADE (C-statistic: 0.595, 95% CI: 0.567-0.622). However, the calibration was good. PRECISE-DAPT showed a higher integrated discrimination improvement (IDI) than PARIS-MB, HAS-BLED, ATRIA, and CRUSADE (P < 0.05) and the best decision curve analysis (DCA). For thromboembolic events, the discrimination of GRACE (C-statistic: 0.636, 95% CI: 0.608-0.662) was higher than CHA2DS2-VASc (C-statistic: 0.612, 95% CI: 0.584-0.639), OPT-CAD (C-statistic: 0.602, 95% CI: 0.574-0.629) and PARIS-CTE (C-statistic: 0.595, 95% CI: 0.567-0.622). The calibration was good. Compared to OPT-CAD and PARIS-CTE, the IDI of the GRACE score slightly improved (P < 0.05). However, NRI analysis showed no significant difference. DCA showed that the clinical practicability of thromboembolic risk scores was similar. CONCLUSIONS: The discrimination and calibration of existing risk scores in predicting 1-year thromboembolic and bleeding events were unsatisfactory in elderly patients with comorbid AF and ACS. PRECISE-DAPT showed higher IDI and DCA than other risk scores in predicting BARC class ≥ 3 bleeding events. The GRACE score showed a slight advantage in predicting thrombotic events.
ABSTRACT
ABSTRACT: This study is a systematic review of characteristics and influencing factors of nonsuicidal self-injury behavior among adolescents with depressive disorder in China. PubMed, CNKI, WanFang Database, and VIP were searched for studies. Meta-analysis was performed using RevMan 5.1 software. Nineteen studies involving 707 subjects were included in the meta-analysis. Age, gender, only child or not, and residence were included in the analysis, of which age ( I2 = 0%, p = 0.42) and residence ( I2 = 0%, p = 0.84) were analyzed by fixed-effects model; gender ( I2 = 75%, p = 0.003) and only child or not ( I2 = 50%, p = 0.140) were analyzed by random-effects model. The evidence shows that, according to the common self-injury mode and location, the nonsuicidal self-injury behavior of young people with depressive disorder aged 15-18 years is paid attention to and guided, so as to achieve early detection and early diagnosis and treatment, and reduce the occurrence of serious harm.
Subject(s)
Depressive Disorder , Self-Injurious Behavior , Child , Humans , Adolescent , Self-Injurious Behavior/epidemiology , China/epidemiology , Databases, Factual , Depressive Disorder/epidemiologyABSTRACT
Genetic variants among individuals have been associated with ineffective control of hypertension. Previous work has shown that hypertension has a polygenic nature, and interactions between these loci have been associated with variations in drug response. Rapid detection of multiple genetic loci with high sensitivity and specificity is needed for the effective implementation of personalized medicine for the treatment of hypertension. Here, we used a cationic conjugated polymer (CCP)-based multistep fluorescence resonance energy transfer (MS-FRET) technique to qualitatively analyze DNA genotypes associated with hypertension in the Chinese population. Assessment of 10 genetic loci using this technique successfully identified known hypertensive risk alleles in a retrospective study of whole-blood samples from 150 patients hospitalized with hypertension. We then applied our detection method in a prospective clinical trial of 100 patients with essential hypertension and found that personalized treatment of patients with hypertension based on results from the MS-FRET technique could effectively improve blood pressure control rate (94.0% versus 54.0%) and shorten the time duration to controlling blood pressure (4.06 ± 2.10 versus 5.82 ± 1.84 days) as compared with conventional treatment. These results suggest that CCP-based MS-FRET genetic variant detection may assist clinicians in rapid and accurate classification of risk in patients with hypertension and improve treatment outcomes.
Subject(s)
Antihypertensive Agents , Hypertension , Humans , Fluorescence Resonance Energy Transfer/methods , Hypertension/drug therapy , Hypertension/genetics , Polymers , Polymorphism, Single Nucleotide/genetics , Prospective Studies , Retrospective StudiesABSTRACT
PURPOSE: To determine the association between ammonium concentration in culture medium and blastocyst development and to assess the influence of increased ammonium concentration on the expression of Bax, Bcl-2 and Oct4. METHODS: A total of 254 cleavage-stage embryos were individually cultured in 30µL G2-plus medium on Day 3, and then culture media samples were collected on Day 5 for ammonium concentration determination immediately after evaluating the embryos morphology. Poor-quality blastocysts (combined score of CC) were used for gene expression analysis. The blastocyst formation rate, good-quality blastocyst rate and relative expression levels of Bax, Bcl-2 and Oct4 were analyzed. RESULTS: Based on receiver operating characteristic curve, the cutoff value of ammonium concentration produced by embryos was 16.07 µmol/L (AUC = 0.722, 95% CI 0.637-0.807; P = 0.000), so all embryos were assigned to two groups according to the cutoff value: normal group (< 16.07 µmol/L) and increased group (≥ 16.07 µmol/L). There was a significant difference in blastocyst formation rate (80.5% vs 59.0%, P < 0.01) between normal group and increased group, as well as for good-quality blastocyst rate (21.0% vs 3.4%, P < 0.01). A significantly higher expression level of Bax (P < 0.05) and considerably lower expression level of Oct4 (P < 0.01) were observed in increased group compared to normal group. CONCLUSION: Our data demonstrated for the first time that increased ammonium concentration in culture medium may promote cellular apoptosis and negatively affect pluripotency of human blastocyst.
Subject(s)
Ammonium Compounds , Humans , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolism , Ammonium Compounds/metabolism , Embryo Culture Techniques , Blastocyst/metabolism , Embryonic Development , Proto-Oncogene Proteins c-bcl-2/metabolism , Apoptosis/geneticsABSTRACT
Zygotes without a pronuclear (0PN) or with one pronuclear (1PN) were defined as abnormal fertilization in conventional in vitro fertilization (IVF). The removal of 0PN and 1PN zygotes from conventional IVF cycles has always been controversial. This study aimed to investigate the chromosomal aneuploidy rates of 0PN- and 1PN-derived blastocysts in conventional IVF cycles and to assess the concordance rate between TE-biopsy PGT-A and miPGT-A. TE biopsies and culture media with blastocoel fluid (CM-BF) samples were whole-genome amplified by multiple annealing and looping-based amplification cycle-based single-cell ChromInst method. Next generation sequencing was performed for comprehensive chromosomal screening on a NextSeq550 sequencer using the NextSeq 500/550 High Output kit v2. The aneuploidy rates of 0PN-derived blastocysts were 19.7% for TE-biopsy PGT-A, and 36.1% for miPGT-A; the concordance rate for ploidy was 77.0%; and the sensitivity and specificity were 83.3% and 75.5%, respectively. The aneuploidy rates of 1PN-derived blastocysts were 37.5% and 37.5% by TE-biopsy PGT-A and miPGT-A, respectively; the concordance rate between TE biopsies and CM-BF samples was 83.3%; and the sensitivity and specificity were 77.8% and 86.7%, respectively. Regarding TE-biopsy PGT-A, there were no significant differences in aneuploidy rates among 0PN-, 1PN- and 2PN-derived blastocysts (PGT-M cycles) (19.7% vs. 37.5% vs. 24.3%, P = 0.226), but the aneuploidy rate of 1PN-derived blastocysts was slightly higher than the other two groups. An increase in aneuploidy rates was observed for 0PN/1PN-derived day 6 blastocysts compared to 0PN/1PN-derived day 5 blastocysts (TE-biopsy PGT-A: 35.7% vs. 19.3%, P = 0.099; miPGT-A: 39.3% vs. 35.1%, P = 0.705). The present study is the first that contributes to understanding the chromosomal aneuploidies in 0PN- and 1PN-derived blastocysts in conventional IVF cycles using TE-biopsy PGT-A and miPGT-A. The clinical application value of 0PN- and 1PN-derived blastocysts in conventional IVF should be assessed using TE-biopsy PGT-A or miPGT-A due to the existence of chromosomal aneuploidies.. In terms of consistency, the miPGT-A using blastocoel fluid enriched culture medium is promising as an alternative to TE-biopsy PGT-A for aneuploidy testing of 0PN- or 1PN-derived blastocysts in conventional IVF.
ABSTRACT
Background and Aims: The association of familial hypercholesterolemia (FH) with risk of cardiovascular events (CVE) and death in different cohorts is controversial. We aimed to assess the risk of CVE and death in patients with FH in different cohorts, including CHD and ACS patients, White and Asian, different diagnostic criteria. Methods: We searched PubMed, MEDLINE, and Web of Science electronic databases through May 2021 to identify cohort studies of CVE and death in patients with FH. Results: We found 18 eligible studies with 1,139,788 participants, including 34,261 patients. There were 31,287 ACS patients, of whom 2,338 were combined with FH. Randomized-effects meta-analysis showed that in patients with FH, relative risk (RR) of CVE and death was 1.87 (95% CI 1.21-2.88), among which CVE was 2.14 (95%CI 1.26-3.64), all-cause of death RR = 1.12 (95% CI 0.89-1.41), and cardiac death RR = 1.03 (95% CI 0.59-1.79). Risk of CVE and death in general population with FH was 2.85 (95% CI 0.72-11.21), hyperlipidemia population RR = 1.59 (95% CI 1.05-2.41), coronary heart disease patients (CHD) RR = 1.46 (95% CI 1.24-1.72), and acute coronary syndrome patients (ACS) RR = 1.71 (95% CI 1.19-2.46). Among ACS patients, the RR of CVE in patients with FH was 1.91 (95% CI 1.55-2.35), the RR of all-cause of death was 1.03 (95% CI 0.80-1.32), and the RR of cardiac death was 1.03 (95% CI 0.59-1.79). The risk of CVE and death in ACS patients with FH in White was 1.69 (95% CI 1.09-2.64) and Asian 1.90 (95% CI 1.31-2.75). RR in patients with Dutch Lipid Network criteria (DLCN) ≥6 vs. <3 points was higher (RR = 2.24, 95% CI 1.69-2.97). RR for long-term follow-up was 1.68 (95% CI 1.09-2.61) and for short-term follow-up was 1.80 (95% CI 1.16-2.78). The results of the overall population were similar, but RR for overall population during a short-term follow-up was 1.49 (95% CI 0.81-2.73). We followed PRISMA checklist to complete meta-analysis. Conclusions: The risk of CVE and death was increased in patients with CHD, especially in patients with ACS. DLCN ≥ 6 points was suggested for clinical diagnosis of FH. The risk of long-term and short-term CVE and death increased in ACS patients with FH. Registration Number: INPLASY2021110010.
ABSTRACT
RATIONALE AND OBJECTIVES: To assess the diagnostic performance of dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) and fluorine-18-fluorodeoxyglucose (18F-FDG) positron-emission tomography/computed tomography (PET/CT) parameters in evaluating the biological behavior of soft tissue tumors. MATERIALS AND METHODS: We retrospectively analyzed DCE-MRI and 18F-FDG PET/CT parameters in 78 patients with pathology-confirmed soft tissue tumors. A total of 78 patients had undergone DCE-MRI examination, while 24 patients with malignant soft tissue tumor had undergone 18F-FDG PET/CT examination. Microvessel density (MVD) and the Ki-67 labeling index (LI) were detected using immunohistochemistry. Differences in parameters (Ktrans, Kep, Ve, MVD, and Ki-67 LI) between benign and malignant tumors were compared. Differences in parameters (Ktrans, Kep, Ve, MVD, and SUVmax) between high- and low-proliferation malignant tumors (grouped by Ki-67 LI) were compared. Correlation of the DCE-MRI and 18F-FDG PET/CT parameters with MVD and Ki-67 LI was analyzed. RESULTS: Only the Ktrans, Kep, MVD, and Ki-67 LI differed significantly between the benign and malignant soft tissue tumors (all p < 0.001). Only Kep (p = 0.033) and SUVmax (p = 0.001) differed significantly between high- and low-proliferation malignant soft tissue tumors. Ktrans, Kep, and SUVmax correlated positively with MVD (r = 0.805, 0.778, 0.730, respectively; all p < 0.001), and with Ki-67 LI (r = 0.721, 0.685, 0.655, respectively; all p < 0.001). CONCLUSION: DCE-MRI and 18F-FDG PET/CT parameters indicate soft tissue tumor biological behavior and can be used to differentiate between benign and malignant soft tissue tumors and between high- and low-proliferation malignant soft tissue tumors.
Subject(s)
Fluorodeoxyglucose F18 , Soft Tissue Neoplasms , Humans , Positron Emission Tomography Computed Tomography/methods , Retrospective Studies , Ki-67 Antigen , Contrast Media , Magnetic Resonance Imaging/methods , Soft Tissue Neoplasms/diagnostic imaging , Positron-Emission Tomography/methodsABSTRACT
BACKGROUND: Up-regulation of aerobic glycolysis has been reported as a characterization of asthma and facilitates airway inflammation. We has been previously reported that short isoform thymic stromal lymphopoietin (sTSLP) could reduce inflammation in asthmatic airway epithelial cells. Here we wanted to investigate whether the inhibition of sTSLP on asthma is related to aerobic glycolysis. METHODS: Asthmatic model was established in challenging Male BALB/c mice and 16-HBE (human bronchial epithelial) cell line with house dust mite (HDM). Indicators of glycolysis were assessed to measure whether involve in sTSLP regulating airway epithelial cells inflammation in asthmatic model in vivo and in vitro. RESULTS: sTSLP decreased inflammation of asthmatic airway and aerobic glycolysis in mice. HDM or long isoform thymic stromal lymphopoietin (lTSLP) promoted HIF-1α expression and aerobic glycolysis by miR-223 to target and inhibit VHL (von Hippel-Lindau) expression 16-HBE. Inhibition of aerobic glycolysis restrained HDM- and lTSLP-induced inflammatory cytokines production. sTSLP along had almost no potential to alter aerobic glycolysis of 16-HBE. But sTSLP decreased LDHA (lactate dehydrogenase A) and LD (Lactic acid) levels in BALF, and HIF-1α and LDHA protein levels in airway epithelial cells of asthma mice model. lTSLP and sTSLP both induced formation of TSLPR and IL-7R receptor complex, and lTSLP obviously facilitated phosphorylation of JAK1, JAK2 and STAT5, while sTSLP induced a little phosphorylation of JAK1 and STAT5. CONCLUSION: We identified a novel mechanism that lTSLP could promote inflammatory cytokines production by miR-223/VHL/HIF-1α pathway to upregulate aerobic glycolysis in airway epithelial cells in asthma. This pathway is suppressed by sTSLP through occupying binding site of lTSLP in TSLPR and IL-7R receptor complex.
Subject(s)
Asthma , Cytokines , Animals , Asthma/metabolism , Cytokines/metabolism , Epithelium/metabolism , Glycolysis , Humans , Inflammation/metabolism , Male , Mice , Protein Isoforms , Thymic Stromal LymphopoietinABSTRACT
PURPOSE: The aim of this study was to determine factors affecting the chromosome imbalance in blastocysts and reproductive outcomes by a comparison between the reciprocal translocation (REC), inversion (INV), and Robertsonian translocation (ROB) carriers. METHODS: Couples with one partner carrying translocation or inversion underwent preimplantation genetic testing for chromosomal structural rearrangement (PGT-SR) cycles, including 215 PGT-SR cycles performed in subsequent 164 frozen-thawed embryo transfer cycles and 61 prenatal diagnoses of fetuses and 59 normal live birth babies. A total of 899 samples were processed by whole-genome amplification followed by next-generation sequencing (NGS). Karyotype and chromosome microarray analyses were used to confirm the PGT results from the amniotic fluid samples. RESULTS: A total of 843 blastocysts from 124 REC, 21 INV, and 35 ROB carriers were diagnosed by PGT-SR. The percentage of unbalanced blastocysts was significantly higher in REC than in INV and ROB carriers (64.31% vs. 28.05% vs. 37.02%). Stratification analysis of female carrier age and gonadotropin doses showed no significant increase in unbalanced chromosomal abnormalities in the three groups. Also, the different breakpoints in chromosomal arms did not affect the rate of unbalanced chromosomes in the embryos. Logistic regression indicated blastocyst quality as a statistically significant risk factor associated with unbalanced chromosomal abnormalities from translocation carriers (P < 0.001). The source of abnormalities in the three groups showed significant differences such that the abnormalities in REC mostly originated from parental translocation but the abnormalities in INV were mainly de novo variations. 164 blastocysts were transferred, and there were no significant differences in the clinical pregnancy rate and miscarriage rate. A total of 59 healthy babies were born, and there were no significant differences in the gender ratio and birth height, except the birth weight of boys between INV and ROB groups (P = 0.02). The results of amniocentesis revealed that more fetuses have normal chromosomal karyotypes than balanced carriers, particularly in the REC group. CONCLUSIONS: Reciprocal translocation carriers have more risk of unbalanced rearrangement, but embryonic chromosome abnormalities of inversion carriers come mainly from de novo variations. This is the first study specifically comparing three different PGT-SRs using the NGS method and evaluating their reproductive outcomes. Our findings will provide the reciprocal translocation, inversion, and Robertsonian translocation carrier couples with more accurate genetic counseling on the reproductive risk of chromosomal imbalance.
Subject(s)
Chromosome Disorders/diagnosis , Chromosomes, Human/chemistry , Fertilization in Vitro/methods , Genetic Testing/methods , High-Throughput Nucleotide Sequencing/methods , Preimplantation Diagnosis/methods , Adult , Chromosome Disorders/genetics , Chromosomes, Human/genetics , Embryo Transfer , Female , Humans , Male , Pregnancy , Pregnancy Outcome , Pregnancy Rate , Retrospective StudiesABSTRACT
OBJECTIVE: We conducted this meta-analysis to assess the efficacy and safety of imiquimod in comparison with other treatments in patients with BCC. METHODS: A comprehensive literature search was performed in the database of PubMed, Embase, and Web of Science. Outcomes of interest included histological/composite clearance rate, success rate, complete response rate, tumor free survival, and adverse events. Pooled risk ratio (RR) with 95% confidence intervals (CIs) using a fixed-effects or random-effects model were determined for each outcome. RESULTS: A total of 13 studies involving 4256 patients were identified. Imiquimod was associated with significantly higher histological clearance rate (RR = 9.28, 95%CI: 5.56, 15.49; P < .001) and composite clearance rate (RR = 34.24, 95%CI: 21.29, 55.06; P = .001). Moreover, imiquimod also significantly increased complete response rate (RR = 3.15, 95%CI: 1.55, 6.38; P = .001) but had no effect in the success rate (RR = 0.98, 95%CI: 0.89, 1.08; P = .727) and probability of tumor-free survival (RR = 1.15, 95%CI: 0.98, 1.35; P = .088), as compared with other treatments. There were more patients in imiquimod group who developed adverse events than in other treatment group (RR = 2.00, 95%CI: 1.39, 2.88; P < .001). CONCLUSION: This study indicated the effects of imiquimod in improving the histological/composite clearance rates as compared with other treatments. However, its treatment-related adverse events also should be noticed. Our findings supported that, imiquimod could be used as the first-choice treatment for BCC.
Subject(s)
Antineoplastic Agents/administration & dosage , Carcinoma, Basal Cell/drug therapy , Imiquimod/administration & dosage , Skin Cream/administration & dosage , Skin Neoplasms/drug therapy , Antineoplastic Agents/adverse effects , Carcinoma, Basal Cell/mortality , Carcinoma, Basal Cell/pathology , Disease-Free Survival , Drug Administration Schedule , Humans , Imiquimod/adverse effects , Neoplasm Grading , Skin Cream/adverse effects , Skin Neoplasms/mortality , Skin Neoplasms/pathologyABSTRACT
PURPOSE: Empty follicle syndrome (EFS) is a complex reproductive disorder characterized by the repeated failure to aspirate oocytes from mature ovarian follicles during in vitro fertilization (IVF). In addition to some cases caused by iatrogenic problems and known genetic factors, there are still many unexplained aspects of EFS. Here, we aimed to assess the clinical and genetic characteristics of two EFS patients. METHODS: We have characterized two primary infertility patients with EFS in a nonconsanguineous family from China. Both the patients presented similar clinical phenotypes, that is a few granulosa cells but no oocytes could be retrieved during repeated cycles with normal follicular development, E2 levels, and bioavailable hCG plasma levels. Abnormal oocytes were obtained once or twice between multiple IVF cycles. We performed Sanger sequencing of the LHCGR and ZP1~ZP4 genes in the patients, and further bioinformatics analysis was performed to identify pathogenic elements in the genes. RESULTS: A novel mutation, c.181C>T (p.Arg61Cys), and a known mutation, c.1169_1176delTTTTCCCA (p.Ile390Thrfs*16), in the ZP1 gene were both identified in patient 2, but no mutations were identified in patient 1. The novel mutation inherited from her mother was absent in the control cohort and the ExAc database. The arginine residue is conserved at this position, and its replacement by cysteine was predicted to be deleterious. In another allele, a paternal frameshift mutation was predicted to introduce premature stop codons, resulting in the deletion of 234 amino acids from the C-terminus of the ZP1 protein. CONCLUSIONS: Our findings presented compound heterozygous mutations in ZP1 associated with EFS and abnormal oocytes and provided further new evidence for the genetic basis of EFS and support for the genetic diagnosis of infertile individuals.
Subject(s)
Genetic Predisposition to Disease , Infertility, Female/genetics , Ovarian Diseases/genetics , Zona Pellucida Glycoproteins/genetics , Adult , China/epidemiology , Female , Humans , Infertility, Female/pathology , Mutation , Oocytes/growth & development , Oocytes/pathology , Ovarian Diseases/pathology , Ovarian Follicle/growth & development , Ovarian Follicle/pathology , Ovulation/genetics , Ovulation Induction/methods , Phenotype , Zona Pellucida/pathologyABSTRACT
PURPOSE: Cleavage of the zygote during human reproduction is a key event of early embryonic development. The genetic events associated with idiopathic embryonic cleavage failure are not certain. Mutations in the tubulin beta 8 class VIII (TUBB8) gene have been reported to be associated with oocyte maturation, fertilization, and developmental arrest. Here, we aimed to assess the clinical and genetic characteristics of complete cleavage failure in fertilized eggs. METHODS: We have characterized a patient with a 9-year history of primary infertility in a non-consanguineous family from China. The patient presented complete cleavage failure in all two-pronuclear (2PN) fertilized oocytes after 2 cycles of in vitro fertilization (IVF). We performed Sanger sequencing of the TUBB8 gene in the patient, and further bioinformatics analysis to identify pathogenesis of gene. RESULTS: A novel homozygous mutation, c.322G > A (p.Glu108Lys), was detected, and this change was absent from 179 control subjects. Glutamic acid is highly conserved at this position, and replacement by lysine was predicted to be repelled by the α-tubulin positive region, disrupting the α-ß tubulin interaction. CONCLUSIONS: Our findings presented a homozygous mutation of TUBB8 associated with complete cleavage failure in fertilized eggs and provided new data for the genotype-phenotype of TUBB8-related diseases.
Subject(s)
Cleavage Stage, Ovum/physiology , Embryonic Development/physiology , Mutation/genetics , Oocytes/physiology , Tubulin/genetics , Adult , China , Female , Fertilization in Vitro/methods , Humans , Infertility, Female/genetics , Zygote/physiologyABSTRACT
PURPOSE: The purpose of this study was to evaluate the effects of mitochondrial supplementation (MS) on early embryonic development and to assess the safety of MS treatments using induced pluripotent stem cells (iPSCs) as the mitochondrial donor. METHODS: In this study, we evaluated the effect of MS on early embryonic development using induced pluripotent stem cells (iPSCs) as the donor. Mouse zygotes were injected with either mitochondria from iPSCs or a vehicle solution. Several parameters were evaluated, including the rates of blastocyst formation and implantation, the weight of E13.5 embryos and placentas, the distribution of the donor mitochondrial DNA (mtDNA), and the pattern of methylation in the differentially methylated regions (DMRs) of the H19 and Snrpn genes. RESULTS: We found that neither the rates of blastocyst formation and implantation nor the weights of E13.5 embryos and placentas were significantly different between the MS and control groups. Additionally, the mtDNA from the iPSC donors could be detected in the muscle tissue of four fetuses and all placentas in the MS group. Finally, the methylation patterns of H19 and Snrpn DMRs remained unchanged by MS. CONCLUSIONS: iPSC-derived mtDNA was directly involved in the process of embryonic development after MS. No adverse effects were seen when using iPSCs as a mitochondrial donor, but it remains to be seen whether this method can improve embryonic development, especially in older mice.
Subject(s)
Embryonic Development/physiology , Induced Pluripotent Stem Cells/cytology , Mitochondria/physiology , Animals , Blastocyst/cytology , DNA Methylation/physiology , DNA, Mitochondrial/genetics , Embryo Implantation/physiology , Female , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Placenta/cytology , PregnancyABSTRACT
Empty follicle syndrome (EFS) is a reproductive disorder in which no oocytes are retrieved during IVF. The existence of genuine EFS (GEFS) is still controversial, and to date, only one missense mutation of Luteinizing Hormone/Choriogonadotropin Receptor (LHCGR) has been reported to be associated with this disease. Here, we describe a GEFS patient in a non-consanguineous family from China. A 27-year-old woman presented with a 5-year history of primary infertility and LH resistance-like ovaries of unequal sizes, but with normal levels of circulating LH. In spite of a satisfactory ovarian reserve and response, no oocytes were retrieved after two cycles of IVF. Her condition did not appear to be failure of the hCG injection. It is more likely to be a genetic cause. A novel homozygous mutation in LHCGR gene, c.1345G>A (p.Ala449Thr), was detected in this patient. Each of her parents is heterozygous for this change, and the change was absent from 407 control subjects. Alanine at this amino acid position was highly conserved and replacement of threonine was predicted to disrupt the third transmembrane helix of the rhodopsin-like G protein-coupled receptor domain. Protein localization studies revealed that a portion of the mutant LHCGR protein molecules was retained intracellularly. Signalling studies demonstrated that this mutation had differing effects on the response of LHCGR to hCG or LH at different concentrations. Specifically, at a concentration <1 IU/ml, the mutant was activated by hCG stimulation but partially resistant to LH stimulation; at a higher concentration (>1 IU/ml), the mutant was activated by both hCG and LH. These data suggest that screening for mutations in the LHCGR gene may assist in the diagnosis of patients with GEFS. The literature describing the relationship between phenotype and genotypes in females is reviewed, and possible aetiologies and treatment options for this disease are proposed based on our and other studies.
Subject(s)
Infertility, Female/genetics , Ovarian Diseases/genetics , Receptors, LH/genetics , Amino Acid Substitution , Female , Genetic Association Studies , Humans , Infertility, Female/pathology , Oocyte Retrieval , Oocytes/growth & development , Protein Structure, Tertiary , Receptors, LH/chemistry , Young AdultABSTRACT
In the modern biological area, the applications of pig as a laboratory model have extensive prospects, such as gene transfer, IVF, SCNT, and xenotransplantation. However, the risk of pathogen transmission by porcine embryos is always a topic to be investigated, especially the viruses related to reproductive failure, for instance, pseudorabies virus, porcine reproductive and respiratory syndrome virus, porcine parvovirus, and porcine circovirus type 2. It should be mentioned that the zona pellucida (ZP) of porcine embryos can be a barrier against the viruses, but certain pathogens may stick to or even pass through the ZP. With intact, free, and damaged ZP, porcine preimplantation embryos are susceptible to these viruses in varying degrees, which may be associated with the virus-specific receptor on embryonic cell membrane. These topics are discussed in the present review.
Subject(s)
Blastocyst/virology , Embryo, Mammalian/virology , Swine/embryology , Viruses/classification , Animals , Zona Pellucida/physiologyABSTRACT
The present aimed to study if porcine circovirus type 2 (PCV2), which adhered to zona pellucida (ZP), was able to enter mature porcine oocytes with intact and damaged ZP. Four groups, including uninfected ZP-intact oocytes (UOZI), uninfected ZP-damaged oocytes (UOZD), PCV2-infected ZP-intact oocytes (POZI), and PCV2-infected ZP-damaged oocytes (POZD) were studied. The oocytes were incubated with 1 mL minimum essential medium, containing 3.1 × 10(8) copies of PCV2 DNA for 1 hour. Mechanical procedure of the insertion by microneedle induced injuries to the ZP of porcine oocytes. At the blastocyst stage, the percentage of PCV2-infected embryos and the ratio of viral antigen-positive cells per embryo were determined by indirect immunofluorescence. To assess the effect of ZP injury on the developmental competence and quality of porcine PCV2-infected oocytes after parthenogenetic activation, blastocyst formation rates and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling staining were analyzed. Moreover, real-time polymerase chain reaction was used to evaluate the expression of genes related to apoptosis and pluripotency at different developmental stages. The results of indirect immunofluorescence showed that only POZD group presented PCV2-infected embryos and viral-positive cells. The blastocyst rate of POZD group dropped down to approximately half of POZI group's (7.1 ± 1.5 vs. 14.5 ± 3.3). At the blastocyst stage, ZP injury increased apoptotic index of PCV2-infected embryos. The relative expression levels of Caspase 3 were higher in POZD group than the ones in POZI group at the two- and four-cell stages (not statistically significant). Compared with the one in POZI group, the ratio of antiapoptotic Bcl-xl gene to proapoptotic Bax gene, an indicator of the ability to resist apoptosis, was lower in POZD group at the one-cell stage, but higher at the two- and four-cell stages. Expression levels of Oct4 and Nanog associated with pluripotency were lower in POZD group than the ones in POZI group at the morula stage (not statistically significant). Noteworthily, the expression of Nanog was significantly lower in POZD group versus POZI group (P < 0.05), whereas relative expression of Oct4 was significantly higher in the former at the blastocyst stage (P < 0.01). In conclusion, PCV2, which attached to ZP, was able to enter mature porcine oocytes with damaged ZP and subsequently reduced the developmental competence and quality of the oocytes after parthenogenetic activation.
Subject(s)
Circovirus/physiology , Oocytes/virology , Parthenogenesis/physiology , Swine/physiology , Animals , Antigens, Viral/genetics , Antigens, Viral/metabolism , Apoptosis , Embryo, Mammalian/virology , Female , Gene Expression Regulation, Developmental/physiology , Gene Expression Regulation, Viral/physiology , In Vitro Oocyte Maturation Techniques/veterinary , Oocytes/physiology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Zona PellucidaABSTRACT
OCT4 is a well-established regulator of pluripotency and nuclear reprogramming. To determine if improving OCT4 abundance can facilitate oocyte-mediated reprogramming in cloned porcine embryos, we artificially increased OCT4 levels by co-incubating donor cells with 50 ng/µl OCT4 plasmid. We observed higher rates of blastocyst formation (P < 0.05) and lower levels of blastocyst apoptosis in nuclear-transfer-derived embryos carrying OCT4-incubated donor nuclei (OCT4-SCNT). The beneficial effect caused by exogenous expression of OCT4 involves epigenetic changes, wherein increased histone acetylation (AcH3K9) appeared in OCT4-SCNT embryos at the one-cell and blastocyst stages and reduced histone methylation (H3K9me2) was observed at the one-cell stage (P < 0.05). There was a transient increase in exogenous OCT4 and an up-regulation of endogenous OCT4 level in OCT4-SCNT embryos (P < 0.05), while the expression pattern of epigenetic enzymes was changed. These modifications were accompanied by an up-regulation of CDX2, whose interaction with OCT4 is instrumental for implantation, and a down-regulation of XIST, a negative indicator of reprogramming (P < 0.05). Taken together, our results support a role for exogenous expression of OCT4 in improving the efficiency of nuclear reprogramming while establishing a convenient and timesaving method to improve nuclear-transfer outcomes.