Appropriate whole genome amplification and pathogenic loci detection can improve the accuracy of preimplantation genetic diagnosis for deletional α-thalassemia

ObjectiveTo improve the accuracy of preimplantation genetic testing (PGT) in deletional α-thalassemia patients.DesignArticle.Patient(s)fifty-two deletional α-thalassemia couples.Intervention(s)Whole genome amplification (WGA), Next-generation sequencing (NGS) and PCR mutation loci detection.Main outcome measuresWGA, Single nucleotide polymorphism (SNP) and PCR mutation loci detection results; Analysis of embryo chromosome copy number variation (CNV).ResultsMultiple Displacement Amplification (MDA) and Multiple Annealing and Looping–Based Amplification Cycles (MALBAC) methods for PGT for deletional α-thalassemia. Blastocyst biopsy samples (n = 253) were obtained from 52 deletional α-thalassemia couples. The results of the comparison of experimental data between groups MALBAC and MDA are as follows: (i) The average allele drop-out (ADO) rate, MALBAC vs. MDA = 2.27% ± 3.57% vs. 0.97% ± 1.4%, P=0.451); (ii) WGA success rate, MALBAC vs. MDA = 98.61% vs. 98.89%, P=0.851; (iii) SNP haplotype success rate, MALBAC vs. MDA = 94.44% vs. 96.68%, P=0.409; (iv) The result of SNP haplotype analysis is consistent with that of Gap-PCR/Sanger sequencing results, MALBAC vs. MDA = 36(36/72, 50%) vs. 151(151/181, 83.43%), P=0; (v) Valid SNP loci, MALBAC vs. MDA = 30 ± 9 vs. 34 ± 10, P=0.02; (vi) The mean CV values, MALBAC vs. MDA = 0.12 ± 0.263 vs. 0.09 ± 0.40, P=0.916; (vii) The average number of raw reads, MALBAC vs. MDA =3244259 ± 999124 vs. 3713146 ± 1028721, P=0; (viii) The coverage of genome (%), MALBAC vs. MDA = 5.02 ± 1.09 vs. 5.55 ± 1.49, P=0.008.ConclusionsOur findings indicate that MDA is superior to MALBAC for PGT of deletional α-thalassemia. Furthermore, SNP haplotype analysis combined with PCR loci detection can improve the accuracy and detection rate of deletional α-thalassemia

Temperature impacts on the growth of hydrogen bubbles during ultrasonic vibration-enhanced hydrogen generation

To improve the hydrogen precipitation performance on the surface of the catalytic layer of the proton exchange membrane (PEM) hydrogen cathode, ultrasonic vibration was employed to accelerate the detachment of hydrogen bubbles on the surface of the catalytic layer. Based on the energy and mechanical analyses of nano and microbubbles, the hydrogen bubble generation mechanism and the effect of temperature on bubble parameters during the evolution process when the ultrasonic field is coupled with the electric field are investigated. The nucleation frequency of the hydrogen bubbles, the relationship between the pressure and temperature and the operating temperature during the generation and detachment of bubbles as well as the detachment radius of bubbles under the action of the ultrasonic field are obtained. The effects of ultrasound and temperature on hydrogen production were verified by visual experiments. The results show that the operating temperature affects the nucleation, growth, and detachment processes of hydrogen bubbles. The effect of temperature on the nucleation frequency of bubbles mainly comes from the Gibbs free energy required for the electrolysis reaction. The bubble radius and growth rate are both related to the temperature to the power of one-third. Ultrasonic waves enhance the separation of hydrogen bubbles from the catalyst surface by acoustic cavitation and impact effects. An increase in the working temperature reduces the activation energy barriers to be overcome for the electrolysis reaction of water, which together with a decrease in the Gibbs free energy and the surface tension coefficient, leads to an increase in the nucleation frequency of the catalytic layer and a decrease in the radius of bubble detachment, and thus improves the hydrogen precipitation performance. Visualization experiments show that in actual PEM hydrogen production, ultrasonic intensification can promote the formation of nucleation sites. The ultrasonic induced fine bubble flow not only has a drag effect on the bubble, but also intensifies the polymerization growth of the bubble due to the impact of the fine bubble flow, thus speeding up the detachment of the bubble, shortening the covering time of the hydrogen bubble on the surface of the catalytic electrode, reducing the activation voltage loss and improve the hydrogen production efficiency of PEM. The experimental results show that when the electrolyte is 60°C, the maximum hydrogen production efficiency of ultrasound is increased by 7.34%, and the average hydrogen production efficiency is increased by 5.83%

A Fuzzy Switching Filter for Removing Impulse Noise

There are many works on the restoration of images corrupted by impulse noise. In[1], Stefan Schulte introduced a fuzzy method to detect the impulse noise. This method has impressive quantitative results for removing the noise.However , when applied to images in which many pixels have the same intensity of the noise, it produces many false-hits. In this paper, we introduce a fuzzy-switching filter based[1] can be applied to more images.NCSP\u270

Effect of different artificial shrinkage methods, when applied before blastocyst vitrification, on perinatal outcomes

Abstract Background In recent years, single blastocyst transfer combined with vitrification has been applied widely, which can maximize the cumulative pregnancy rate in per oocyte retrieval cycles and minimize the multiple pregnancy rate. Thus, the guarantee for these is the effectiveness of vitrified blastocyst. Studies has shown that AS of the blastocoel cavity prior to vitrification can reduce injuries, increase the thawed blastocyst survival rate and implantation rate. Several AS methods have been established. However, only a few studies have compared the effectiveness and safety of these AS methods. In this study, we aimed to compare the clinical outcomes and neonatal outcomes in FET cycles with single blastocyst that were artificially shrunk before vitrification by either LAS or MNAS method. Methods A retrospective comparative study of FET cycles in infertile patients which were at our clinic between January 2013 and December 2014. These FET cycles were divided into two groups by the shrinking methods used before vitrification and the clinical and neonatal outcomes were assessed. Results There were no statistically differences in blastocyst survival rates (95.40% vs 94.05%, P > 0.05) between the LAS and MNAS groups. However, compared with MNAS, LAS improved the warmed blastocyst implantation/clinical pregnancy rate (60.82% vs 54.37%, P  0.05). Conclusions No significant differences in neonatal outcomes were observed, while in clinical outcomes, LAS improved the warmed blastocyst implantation/clinical pregnancy rate and live birth rate markedly, there was also an increased risk of monozygotic twin pregnancies

Transcriptome Analysis of Intermittent Light Induced Early Bolting in Flowering Chinese Cabbage

In flowering Chinese cabbage, early booting is one of the most important characteristics that is linked with quality and production. Through fixed light intensity (280 μmol·m−2·s−1) and fixed intermittent lighting in flowering Chinese cabbage, there was early bolting, bud emergence, and flowering. Moreover, the aboveground fresh weight, blade area, dry weight of blade, and quantification of the leaves in flowering Chinese cabbage were significantly reduced, while the thickness of tillers, tillers height, dry weight of tillers, and tillers weight were significantly increased. The chlorophyll contents and soil–plant analysis and development (SPAD) value decreased in the early stage and increased in the later stage. The nitrate content decreased, while the photosynthetic rate, vitamin C content, soluble sugar content, soluble protein content, phenolic content, and flavonoid content increased, and mineral elements also accumulated. In order to explore the mechanism of intermittent light promoting the early bolting and flowering of ‘49d’ flowering Chinese cabbage, this study analyzed the transcriptional regulation from a global perspective using RNA sequencing. A total of 17,086 differentially expressed genes (DEGs) were obtained and 396 DEGs were selected that were closely related to early bolting. These DEGs were mainly involved in pollen wall assembly and plant circadian rhythm pathways, light action (34 DEGs), hormone biosynthesis and regulation (26 DEGs), development (21 DEGs), and carbohydrate synthesis and transport (6 DEGs). Three hub genes with the highest connectivity were identified through weighted gene co-expression network analysis (WGCNA): BrRVE, BrLHY, and BrRVE1. It is speculated that they may be involved in the intermittent light regulation of early bolting in flowering Chinese cabbage. In conclusion, intermittent light can be used as a useful tool to regulate plant growth structure, increase planting density, enhance photosynthesis, increase mineral accumulation, accelerate growth, and shorten the breeding cycle

A new insight into male fertility preservation for patients with completely immotile spermatozoa

Abstract Background Sperm cryopreservation is the most effective method to preserve male fertility but this is normally used for motile spermatozoa. Thus, only motile spermatozoa are used for cryopreservation in most reproductive medicine centers worldwide. The immotile spermatozoa from some problematic patients are usually discarded, resulting in a missed opportunity of sterility cryopreservation for future assisted reproductive treatments. Many studies have shown that successful fertilization can be obtained after selection of viable sperm from the completely immotile spermatozoa before ICSI. Whether the completely immotile spermatozoa are worth of freezing has not been realized The aim of this study is to explore the clinical value of cryopreservation of immotile spermatozoa. Methods Completely immotile spermatozoa were collected and frozen, and subsequently viable but immotile frozen-thawed spermatozoa were selected by laser plus for ICSI. Main outcomes included spermatozoa survival index, fertilization rate and good quality embryo rate. Results After identification by laser, the fresh samples of spermatozoa presented with a mean survival rate of 54.86% and 26.05%, and this was reduced to 44.13% and 18.13% in frozen-thawed spermatozoa samples, which showed a frozen-thawed spermatozoa survival index of 0.80 and 0.70 in the testicular and ejaculate sperm, respectively. There were no statistically differences in fertilization rate (80% vs80.51%, 75.00% vs 81.48%), cleavage rate (95.45% vs 98.95%, 100.00% vs 95.45%) and good quality embryo rate (40.48% vs 52.13%, 33.33%vs38.10%) between the frozen-thawed immotile spermatozoa group and the routine fresh immotile spermatozoa ICSI group in both testicular and ejaculate sperm, respectively. Conclusions The results of the study show that completely immotile spermatozoa can be frozen in order to preserve male fertility as long as viable spermatozoa are present. This procedure provides a further possibility for fertility preservation for patients with completely immotile spermatozoa

Table_2_Appropriate whole genome amplification and pathogenic loci detection can improve the accuracy of preimplantation genetic diagnosis for deletional α-thalassemia.docx

ObjectiveTo improve the accuracy of preimplantation genetic testing (PGT) in deletional α-thalassemia patients.DesignArticle.Patient(s)fifty-two deletional α-thalassemia couples.Intervention(s)Whole genome amplification (WGA), Next-generation sequencing (NGS) and PCR mutation loci detection.Main outcome measuresWGA, Single nucleotide polymorphism (SNP) and PCR mutation loci detection results; Analysis of embryo chromosome copy number variation (CNV).ResultsMultiple Displacement Amplification (MDA) and Multiple Annealing and Looping–Based Amplification Cycles (MALBAC) methods for PGT for deletional α-thalassemia. Blastocyst biopsy samples (n = 253) were obtained from 52 deletional α-thalassemia couples. The results of the comparison of experimental data between groups MALBAC and MDA are as follows: (i) The average allele drop-out (ADO) rate, MALBAC vs. MDA = 2.27% ± 3.57% vs. 0.97% ± 1.4%, P=0.451); (ii) WGA success rate, MALBAC vs. MDA = 98.61% vs. 98.89%, P=0.851; (iii) SNP haplotype success rate, MALBAC vs. MDA = 94.44% vs. 96.68%, P=0.409; (iv) The result of SNP haplotype analysis is consistent with that of Gap-PCR/Sanger sequencing results, MALBAC vs. MDA = 36(36/72, 50%) vs. 151(151/181, 83.43%), P=0; (v) Valid SNP loci, MALBAC vs. MDA = 30 ± 9 vs. 34 ± 10, P=0.02; (vi) The mean CV values, MALBAC vs. MDA = 0.12 ± 0.263 vs. 0.09 ± 0.40, P=0.916; (vii) The average number of raw reads, MALBAC vs. MDA =3244259 ± 999124 vs. 3713146 ± 1028721, P=0; (viii) The coverage of genome (%), MALBAC vs. MDA = 5.02 ± 1.09 vs. 5.55 ± 1.49, P=0.008.ConclusionsOur findings indicate that MDA is superior to MALBAC for PGT of deletional α-thalassemia. Furthermore, SNP haplotype analysis combined with PCR loci detection can improve the accuracy and detection rate of deletional α-thalassemia.</p

Table_1_Appropriate whole genome amplification and pathogenic loci detection can improve the accuracy of preimplantation genetic diagnosis for deletional α-thalassemia.xlsx

ObjectiveTo improve the accuracy of preimplantation genetic testing (PGT) in deletional α-thalassemia patients.DesignArticle.Patient(s)fifty-two deletional α-thalassemia couples.Intervention(s)Whole genome amplification (WGA), Next-generation sequencing (NGS) and PCR mutation loci detection.Main outcome measuresWGA, Single nucleotide polymorphism (SNP) and PCR mutation loci detection results; Analysis of embryo chromosome copy number variation (CNV).ResultsMultiple Displacement Amplification (MDA) and Multiple Annealing and Looping–Based Amplification Cycles (MALBAC) methods for PGT for deletional α-thalassemia. Blastocyst biopsy samples (n = 253) were obtained from 52 deletional α-thalassemia couples. The results of the comparison of experimental data between groups MALBAC and MDA are as follows: (i) The average allele drop-out (ADO) rate, MALBAC vs. MDA = 2.27% ± 3.57% vs. 0.97% ± 1.4%, P=0.451); (ii) WGA success rate, MALBAC vs. MDA = 98.61% vs. 98.89%, P=0.851; (iii) SNP haplotype success rate, MALBAC vs. MDA = 94.44% vs. 96.68%, P=0.409; (iv) The result of SNP haplotype analysis is consistent with that of Gap-PCR/Sanger sequencing results, MALBAC vs. MDA = 36(36/72, 50%) vs. 151(151/181, 83.43%), P=0; (v) Valid SNP loci, MALBAC vs. MDA = 30 ± 9 vs. 34 ± 10, P=0.02; (vi) The mean CV values, MALBAC vs. MDA = 0.12 ± 0.263 vs. 0.09 ± 0.40, P=0.916; (vii) The average number of raw reads, MALBAC vs. MDA =3244259 ± 999124 vs. 3713146 ± 1028721, P=0; (viii) The coverage of genome (%), MALBAC vs. MDA = 5.02 ± 1.09 vs. 5.55 ± 1.49, P=0.008.ConclusionsOur findings indicate that MDA is superior to MALBAC for PGT of deletional α-thalassemia. Furthermore, SNP haplotype analysis combined with PCR loci detection can improve the accuracy and detection rate of deletional α-thalassemia.</p